Hepatocellular carcinoma (HCC) is a common liver cancer-causing the high rate of mortality worldwide. The present study evaluates the possible protective role of lycopene during diethylnitrosamine (DEN)-induced HCC in rats for 16 weeks. A patch of 60 male rats was divided according to the required treatment into 6 groups. DEN was given to rats in drinking water (100 mg/L) to induce HCC. In the present study. The groups supplied with lycopene were administered with lycopene at 5, 10, and 20 mg/kg BW/orally/daily during the experiment. The DEN alone administered group induced alterations in all the investigated parameters. The lycopene pretreated DEN groups showed significant ameliorations in a dose-dependent manner against the deleterious alterations of DEN in the biological markers, oxidative stress, liver function, lipid profile, hematological parameters, serum inflammatory markers (TNF-α and IL-6), immunohistochemical markers, caspase-3 liver content and histopathological changes, where the lycopene dose of 20 mg/kg showed better ameliorative effects. In conclusion, the oral administration of lycopene provides considerable protective effects against DEN-induced HCC which suggested the potential efficacy of lycopene as an additional chemopreventive agent in the treatment of HCC.
Hepatocellular carcinoma (HCC) is a common important form of liver cancer in both incidence and high rate of mortality worldwide 1, 2.
Diethylnitrosamine (DEN) is a chemical used to induce HCC as previously reported 3, 4. When administered in the drinking water, DEN induces liver tumors in rats through induction of oxidative stress in the liver tissue and other organs that causes generation of free radicals as reactive oxygen species (ROS), which initiate peroxidative damage in the liver tissue 5.
The treatment of experimental animals with plant antioxidants like lycopene for a long period could augment the cellular endogenous antioxidants and defend against oxidative stress 6, 7. Lycopene is a red phytochemical pigment found in tomatoes that is believed to exert anti-inflammatory, antioxidant and hepatoprotective properties as well as anticancer activity 8, 9. The lycopene structure contains many double bonds that help in the scavenging activity against oxidative stress that causes tissue injuries 10. In addition, lycopene can reverse the loss of antioxidant enzymes induced by exposure to some toxins revealing its protective and antioxidant roles against many toxins 11, 12.
The present study aims to investigate the ameliorative effects of lycopene on hepatocarcinogenesis in diethylnitrosamine-induced male rats including oxidative stress, some biochemical, hematological, serum inflammatory markers (TNF-α and IL-6), immunohistochemical parameters and caspase-3 content as well as histopathological alterations with a view to its possible applications in the clinical field.
The present study used 60 male Wistar albino rats (180-200 g), divided into 6 groups. The animals were obtained from El-Nile Company (Egypt) and were housed in the animal facility of Faculty of Pharmacy (Boys), Al-Azhar University, Nasr-City, Cairo, Egypt. They were supplied with standard food composed of 20% casein, 15% corn oil, 55% corn starch, 5% salt mixture, and 5% vitaminized starch (Egyptian Company of Oils and Soap, Kafr-Elzayat, Egypt), and DEN water or tap water ad libitum. The animals were kept in standard conditions including room temperature (22°C±2°C) and relative humidity (55±3) with a 12-h light/dark cycle.
2.2. Ethical ApprovalAll rats were conducted following the ethical guidelines and complied with the guide for the care and use of laboratory animals. The study was approved and conducted under the supervision of the independent ethics committee of the Faculty of Pharmacy (Boys), Al-Azhar University, Nasr-City, Cairo, Egypt.
2.3. ChemicalsWe used chemicals of analytical grade supplied from standard commercial suppliers.
Lycopene Redivivo (Lot ECBJ9005V, 36275-5G, Sigma Aldrich, St. Louis, USA) (5 g) was mixed in corn oil with vigorous stirring to the desired concentrations of lycopene/ml 13. Lycopene was administered to the rats via oral route at doses of 5, 10, and 20 mg/kg BW/daily/orally.
The used doses of lycopene and the resulted control-like effects of corn oil alone treatments were verified by a pilot study conducted in our lab (data not shown) following the method of Rao and Shen 14.
The DEN (Sigma Aldrich, St. Louis, USA) was prepared in dark bottles as a fresh solution (100 mg/L) and supplied to rats to induce hepatocarcinogenesis following a previous method 3.
2.4. Experimental DesignThe rats were divided into six groups (10 rats each) as follows:
Group I (Control): Normal rats supplied with food and water for16 weeks.
Group II (Lycopene): Rats of this group were administered with freshly prepared lycopene at a dose of 20 mg/kg BW/daily/orally for the experimental period.
Group III (DEN-treated group): Rats were given DEN solution (100 mg/L) for 8 weeks to induce liver tumor followed by 8 weeks of tap water without DEN to allow the tumor to grow.
Group IV (Lycopene 5 mg-DEN-treated group): Rats of this group were pretreated with freshly prepared lycopene (5 mg/kg BW/daily/orally for 2 weeks pretreatment period and 16 weeks during the experiment) and administered by DEN as the same schedule mentioned in group III.
Group V (Lycopene 10 mg-DEN-treated group): Rats of this group were pretreated with freshly prepared lycopene (10 mg/kg BW/daily/orally for 2 weeks pretreatment period and 16 weeks during the experiment) and administered by DEN as the same schedule mentioned in group III.
Group VI (Lycopene 20 mg-DEN-treated group): Rats of this group were pretreated with freshly prepared lycopene (20 mg/kg BW/daily/orally for 2 weeks pretreatment period and 16 weeks during the experiment) and administered by DEN as the same schedule mentioned in group III.
2.5. Serum and Liver Tissue SamplingSamples of blood and liver tissues were collected from anesthetized animals after 16 weeks. To prepare serum samples, blood was drained from the orbital plexus of the eye using capillary puncture, put into plain tubes and centrifuged (4000 rpm, 15 min). The prepared serum samples were preserved at -20ºC until future analysis. The liver samples were obtained from dissected animals, washed with isotonic saline, dried, and weighed immediately.
2.6. Estimation of Physical CharacteristicsWe estimate the relative liver mass, the percentage of body mass gain, the body mass gain%, the percentage of nodule incidence, and the nodular size.
2.7. Biochemical StudyEach liver tissue was homogenized in Tris-lysis buffer. The obtained homogenate samples were centrifuged (3000 rpm, 15 min) and then separated into aliquots and stored at -80°C until used.
The oxidative stress parameters (lipid peroxidation products, TBARS; catalase, CAT; superoxide dismutase, SOD; hepatic reduced glutathione, GSH) were determined using readymade kits (Biodiagnostic Co. Cairo, Egypt).
The serum liver function parameters (aspartate transaminase, AST; alanine transaminase ALT; alkaline phosphatase, ALP; total bilirubin, TBIL; total protein, TP; albumin as well as calculation of globulin and A/G ratio) and lipid profile (triglycerides, TG; total cholesterol, TC; high-density lipoprotein cholesterol, HDL-C; low-density lipoprotein cholesterol, LDL-C) were estimated using readymade kits (Elitech Co., France).
2.8. Estimation of Liver Caspase-3 ContentThe caspase-3 content in liver tissues was evaluated using ELISA kits (CUSABIO, Fannin St., Houston, USA) using methods outlined in the diagnostic kit.
2.9. Hematological MeasurementsThe hematological parameters (Red blood cell, RBC count; hemoglobin, Hb concentration; hematocrit, Hct percentage; platelet count; white blood cell, WBC count) were measured in the blood samples (CBC analyzer, sk9000, U.S).
2.10. Inflammatory MarkersThe tumor necrosis factor-alpha (TNF-α) was estimated in the blood serum of rats by ELISA technique using a kit purchased from BT LAB, Jiaxing, Zhejiang, China, according to the manufacturer’s instructions provided with the TNF-α assay kit (Catalog No: E0764Ra). The interleukin-6 (IL-6) was estimated in the blood serum of rats by ELISA technique using a kit purchased from BT LAB, Jiaxing, Zhejiang, China, according to the manufacturer’s instructions provided with the IL-6 assay kit (Catalog No: E0135Ra).
2.11. Immunohistochemical AssayThe Avidin-Biotin immunoperoxidase complex technique (ABC) 15 was used in the present study to determine the expression of cytokines (TNF-α, IL-6) and α-fetoprotein tumor markers (Zhong Shan Golden Bridge Biological Technology Co., Ltd., Beijing, China). The colored images were processed and the relative expression level (% control) was quantitated by measuring the color intensity value using ImageJ 1.46r software (Wayne Rasband, National Institutes of Health, Bethesda, USA) to produce maps showing only the biomarker's stained areas.
2.12. The Histopathological StudyThe liver tissue samples were fixed, processed, sectioned, and stained according to the methods of Bancroft and Gamble 16.
2.13. Statistical AnalysisData are presented as mean ± SEM. Multiple comparisons were performed using one-way analysis of variance (ANOVA) followed by Tukey–Kramer for post-hoc analysis. Statistical significance was accepted at a level of p<0.05, as appropriate. Data analysis was performed using GraphPad Instat software (version 3.06). Graphs were sketched using GraphPad Prism (ISI® software, USA, version 5).
The lycopene alone, lycopene 10 mg-DEN, and lycopene 20 mg-DEN treated groups showed insignificant changes in body mass gain, body mass gain%, liver mass & the relative liver mass when compared to control group. On the contrary, the DEN alone treated group and the lycopene 5 mg-DEN-treated group recorded significantly reduced body mass gain and body mass gain % in contrast to a significantly increased (p<0.05) liver mass and the relative liver mass as compared to the control group.
However, the lycopene 10 mg-DEN and lycopene 20 mg-DEN treated groups showed significantly increased body mass gain, body mass gain%, in contrast to significantly reduced (p<0.05) liver mass and relative liver mass when compared to the group treated with DEN alone.
Moreover, the lycopene 10 mg-DEN and lycopene 20 mg-DEN treated groups (compared to the lycopene 5 mg-DEN treated group) recorded significantly increased body mass gain and body mass gain%, in contrast to significantly reduced liver mass and the relative liver mass.
3.2. The Effects of Lycopene on the Morphometric Parameters of the Liver Tissues of Different Groups at the End of 16 Weeks are Represented in Table 1 and Figure 1The morphometric parameters of the liver tissues of the control and lycopene alone treated group revealed a smooth liver with a normal appearance at the end of 16 weeks. On the other hand, the morphology of livers in the DEN, lycopene 5 mg-DEN, lycopene 10 mg-DEN, and lycopene 20 mg-DEN treated groups showed enlarged liver with several foci and nodules of HCC.
There was a 100% incidence of nodules in DEN, lycopene 5 mg-DEN, lycopene 10 mg-DEN, and lycopene 20 mg-DEN treated groups. All lycopene DEN treated groups recorded significantly reduced total nodules and nodules of all sizes when compared to the group treated with DEN alone. Additionally, the lycopene 20 mg-DEN-treated group recorded significantly reduced total nodules and nodules of all sizes when compared to the lycopene 5 mg-DEN or the lycopene 10 mg-DEN-treated groups. Also, the lycopene 10 mg-DEN treated group recorded significantly reduced total nodules and nodules of all sizes when compared to the lycopene 5 mg-DEN treated group.
3.3. The Effects of Lycopene on the Oxidative Stress in the Liver Tissues of Different Groups at the End of 16 Weeks are Represented in Table 2The lycopene alone treated group showed insignificant changes in the liver oxidative stress parameters of GSH, SOD, CAT, and TBARS when compared to the control group. On the contrary, the DEN alone, lycopene 5 mg-DEN, lycopene 10 mg-DEN, and lycopene 20 mg-DEN treated groups recorded significantly increased TBARS, in contrast to significantly reduced SOD, CAT, and GSH when compared to the control group.
Moreover, all lycopene DEN-treated groups showed significantly reduced TBARS in the liver and significantly increased levels of GSH, SOD, and CAT in the liver when compared to the group treated with DEN alone. Also, the lycopene 20 mg-DEN-treated group (compared to the lycopene 5 mg-DEN or the lycopene 10 mg-DEN-treated groups) and the lycopene 10 mg-DEN treated group (compared to lycopene the 5 mg-DEN treated group) recorded significantly reduced TBARS and significantly increased levels of SOD, CAT, and GSH in the liver.
3.4. The Effects of Lycopene on the Tissue Caspase-3 content Apoptotic Marker in the Liver Tissue of Different Groups at the End of 16 Weeks Are Represented in Figure 2The lycopene alone treated group showed insignificant changes in the caspase-3 content in the liver when compared to the control group. On the contrary, the DEN and all lycopene DEN treated groups recorded significantly increased liver caspase-3 content when compared to the control group.
Additionally, all lycopene DEN-treated groups recorded significantly decreased liver caspase-3 content when compared to the group treated with DEN alone. Moreover, the lycopene 20 mg-DEN treated group (compared to the lycopene 5 mg-DEN or the lycopene 10 mg-DEN treated groups) and the lycopene 10 mg-DEN-treated group (compared to the lycopene 5 mg-DEN-treated group) recorded significantly reduced liver caspase-3 content in the liver.
The lycopene alone treated group showed insignificant changes in serum liver function parameters (AST, ALT, ALP, TBIL, TP, globulin, albumin, and A/G ratio) when compared to the control group. On the contrary, the DEN, lycopene the 5 mg-DEN, the lycopene 10 mg-DEN, and the lycopene 20 mg-DEN treated groups recorded significantly increased levels of serum AST, ALT, ALP, TBIL, TP, and globulin in contrast to significantly reduced levels of serum albumin and A/G ratio when compared to the control group.
In addition, all lycopene DEN-treated groups recorded significantly reduced levels of serum AST, ALT, ALP, TBIL, TP, and globulin in contrast to significantly increased levels of serum albumin and A/G ratio when compared to the group treated with DEN alone. Also, the lycopene 20 mg-DEN treated group (compared to the lycopene 5 mg-DEN-treated or the lycopene 10 mg-DEN-treated groups) and the lycopene 10 mg-DEN treated group (compared to the lycopene 5 mg-DEN-treated group) recorded significantly reduced levels of serum AST, ALT, ALP, TBIL, globulin, and TP, in contrast, to significantly increased serum albumin and A/G ratio.
3.6. The Effects of Lycopene on the Serum Lipid Parameters and Their Risk Ratios of Different Groups at the End of 16 Weeks Are Represented in Table 4The lycopene alone treated group was similar to the control group in serum lipid parameters and their risk ratios. On the contrary, the DEN and all lycopene DEN-treated groups recorded significantly increased values of serum TG, TC, LDL-C, as well as their risks, in contrast, to significantly reduced values of serum HDL-C when compared to the control group.
Moreover, all lycopene DEN treated groups showed significantly reduced levels of serum TG, TC, LDL-C, and their risks in contrast to significantly increased levels of HDL-C when compared to the group treated with DEN alone. In addition, the lycopene 20 mg-DEN treated group (compared to the lycopene 5 mg-DEN or the lycopene 10 mg-DEN-treated groups) and the lycopene 10 mg-DEN-treated group (compared to the lycopene 5 mg-DEN treated group) recorded significantly reduced levels of serum TG, TC, LDL-C, and their risks in contrast to a significantly increased level of serum HDL-C.
The lycopene alone treated group was similar to the control in the measured hematological parameters. On the other hand, the DEN, the lycopene 5 mg-DEN, the lycopene 10 mg-DEN, and the lycopene 20 mg-DEN treated groups recorded significantly reduced RBC count, Hb concentration, Hct percentage, platelet count, and WBC count when compared to the control group.
The lycopene 5 mg-DEN, the lycopene 10 mg-DEN, and the lycopene 20 mg-DEN treated groups showed significant increase in all measured hematological parameters when compared to the group treated with DEN alone. Moreover, the lycopene 20 mg-DEN treated group (compared to the lycopene 5 mg-DEN or the lycopene 10 mg-DEN-treated groups) and the lycopene 10 mg-DEN-treated group (compared to the lycopene 5 mg-DEN treated groups) recorded significant increase in all measured hematological parameters.
3.8. The Effects of Lycopene on the Immunohistochemical Localization of the Inflammatory Cytokines (TNF-α, IL-6) and α-fetoprotein Tumor Marker Protein Expressions in the Liver Tissue of Different Groups at the End of 16 Weeks Are Represented in Figure 3 – Figure 5The normal control group and the lycopene alone treated group showed a weak expression of TNF-α, IL-6, and α-fetoprotein. On the other hand, the periportal and peri-venular areas of hepatocytes in rat livers subjected to DEN and all lycopene DEN treated groups revealed diffuse upregulation of cytoplasmic reactivity of TNF-α, IL-6, and α-fetoprotein expressions. Interestingly, all lycopene DEN-treated groups exhibited decreased expression of TNF-α, IL-6, and α-fetoprotein compared with DEN only treated group.
Semi-quantitative analysis of immunohistochemical staining expressed as relative intensity of expression of TNF-α, IL-6, and α-fetoprotein protein revealed that DEN group significantly (p<0.05) increased the hepatic expression of TNF-α, IL-6, and α-fetoprotein compared with normal control group. In contrast, the expression of TNF-α, IL-6, and α-fetoprotein in all lycopene DEN-treated groups were significantly (p<0.05) downregulated compared with the group treated with DEN alone. Moreover, the lycopene 20 mg-DEN treated group (compared to the lycopene 5 mg-DEN or the lycopene 10 mg-DEN treated groups) and the lycopene 10 mg-DEN-treated group (compared to the lycopene 5 mg-DEN-treated group) recorded significantly (p<0.05) downregulated TNF-α, IL-6, and α-fetoprotein expressions in the liver.
The lycopene alone treated group showed insignificant changes in the TNF-α and IL-6 serum levels in when compared to the control group. On the contrary, the DEN alone, the lycopene 5 mg-DEN, and the lycopene 10 mg-DEN groups recorded significant increase in the TNF-α and IL-6 serum levels when compared to the control group.
In addition, the lycopene 20 mg-DEN-treated group showed significant decrease in the TNF-α and IL-6 serum levels when compared to the DEN alone group. Finally, there was no significant change between the lycopene 5 mg-DEN, and the lycopene 10 mg-DEN treated groups in the TNF-α and IL-6 serum levels.
3.10. The Effects of Lycopene on the Liver Tissue Histopathology of Different Groups at the End of 16 Weeks Are Represented in Table 7 and Figure 6The histological morphology of the livers showed the normal liver tissue in the control and lycopene only treated groups. The DEN alone treated group and the groups treated with lycopene and DEN showed severe necrotic changes to the character with karyopyknosis, acute cellular swelling in hepatocytes, karyomegaly, apoptosis, sinusoidal and portal congestion, periportal cell infiltration, intra-, and extracellular bile pigment accumulations, and Kupffer cell hyperplasia.
The lycopene 5 mg-DEN, the lycopene 10 mg-DEN, and the lycopene 20 mg-DEN treated groups showed ameliorative morphological effects when compared to the group treated with DEN alone.
The HCC occurs predominantly in patients with chronic liver disease and cirrhosis 17. DEN can induce liver cancer in experimental animals. The model used in the present study was described previously 3. This model could mimic human HCC and allow studying hepatocarcinogenesis and screening of the potential anti-tumor agents on various phases of liver cancer. A tumor induction begins with cellular transformation, growth of cells (hyperproliferation), formation of nodules, and metastatic lesions 18, 19.
The present study recorded body weight loss and an increase in liver mass as the major characteristics of HCC that agree with 20. The present study also recorded significantly reduced body mass gain and significantly increased liver mass and relative liver mass as compared to the control which might be due to the development of liver tumors 2, 21. The formation of nodules is a good sign of liver cancer and there is a correlation between the size of the hyperplastic nodules and the occurrence of HCC 3, 22 as evidenced by the present data.
The present results reveal that the supplementation of DEN-treated rats with Lycopene cause significant amelioration in all physical and morphometric characters in a dose-dependent manner (Lycopene dose of 20 mg was the best treatment) as compared to DEN alone treated group. That agrees with a previous study 9 who observed that lycopene decreased liver preneoplastic foci in DEN-treated rats. In addition, some studies 23, 24 have demonstrated that in DEN-induced experimental hepatocarcinogenesis, lycopene inhibits the development of hepatocellular carcinoma and provide a protective effect by inhibiting proliferation by interrupting intercellular communication by affecting cell surface receptors, especially in cancer cells. Additional studies show that pretreatment with lycopene is effective against hepatic tumor induction, growth, and also lycopene is effective when administered in combination with toxic substances 22, 25.
The liver is a common target for several toxicants 26. The present study recorded significantly increased TBARS, in contrast to significantly reduced GSH, SOD, and CAT in all DEN treated groups when compared to the control group. DEN metabolism occurs in the liver that causes oxidative stress, generates ROS, and induced carcinogenic effects 27. The oxidative stress state injures tissues and damages the biological systems as well as cell membranes leading to carcinogenesis steps (i.e. Initiation, promotion, and progression) 3, 9. The cells in biological systems contain endogenous natural enzymatic antioxidants like SOD and CAT and non-enzymatic antioxidants mainly GSH used in defenses against oxidative stress 28. The present results are consistent with previous work that reported that the treatment of rats with DEN leads to depletion of the previously mentioned endogenous antioxidants 3.
The GSH defends against oxidative stress by maintaining the normal reduced state of cells 29. GSH can scavenge ROS and other radicals by conjugation with GPx and GST, so it is used as a marker for evaluating the anticarcinogenic potential, and counteracting a variety of carcinogens 22, 30. The depletion in GSH level after DEN administration may be due to inhibition of GSH synthetase or due to its diffusion through damaged cell membranes 31.
The SOD is the only enzyme known to utilize free radicals as a substrate and dismutase the superoxide radicals to hydrogen peroxide and oxygen molecules. However, CAT is a supporting enzyme that transforms hydrogen peroxide to water thereby protecting against ROS, and could be used as an index of increased hydrogen peroxide production 32, 33. The decrease in CAT activity in response to the overproduction of H2O2 might be interpreted by the prospect of oxidative alteration of proteins which can have a substantial physiological impact 3, 34.
The current study also recorded that the supplementation of DEN treated rats with lycopene causes significantly reduced liver TBARS, in contrast, to significantly increased liver SOD, CAT, GSH antioxidants in a dose-dependent manner (lycopene dose of 20 mg was the best treatment) as compared to the group treated with DEN alone. This indicates that lycopene has potent antioxidant effects as it can scavenge free radicals and prevent lipid peroxidation 12, 35. Our data are consistent with previous studies which recorded that the dietary supplement of watermelon juice (a rich natural source of lycopene) can inhibit lipid peroxidation in the liver and effectively protect against hepatocellular injury induced by toxicity 6, 36. In addition, similar researches indicate the ability of lycopene to protect the liver against oxidative damage 11, 37, 38, 39.
The increased caspase-3 content in the liver tissue indicates an apoptosis-induction process 40. Caspase-3 is an important apoptotic marker enzyme in the apoptosis process. It acts as an executioner enzyme that altered many types of proteins in the cell and promotes apoptosis 41, 42. The present results record significantly increased caspase-3 content in the liver of all groups treated with DEN compared with the control which indicates the induction of apoptosis.
The lycopene DEN treated groups in the present study recorded significantly inhibited tissue caspase-3 content in a dose-dependent manner (lycopene dose of 20 mg was the best treatment) which reveals the anti-apoptotic property of lycopene. These data are in approval with Hassan et al. 7 who found that caspase-3 content is elevated in DEN-induced hepatocellular carcinoma in rats. On the other hand, supplementation of animals with lycopene led to a reduction in the liver caspase-3 content in a dose-dependent manner in comparison with rat groups treated with DEN alone and did not receive the previous supplementation suggesting the anti-apoptotic and protective effects of lycopene as consistent with previous studies 43, 44.
The hepato-specific enzymes were increased at hepatocellular damage 3. The liver function parameters in the present study recorded significantly increased liver enzymes, serum total protein, and globulin concentration in contrast to significantly reduced albumin & albumin-globulin ratio in all DEN treated groups when compared to the control. The measured liver function parameters in the present study are generally accepted as an index of liver damage 4, 45. The increase in serum AST & ALT indicates the impairment of the cell membranes of the liver and leakage of these enzymes to the blood 3. In addition, the elevation in ALP activity and TBIL may be due to the large bile duct obstruction, intrahepatic cholestasis, or infiltrative release from the tissues to the blood 46. The serum proteins are synthesized in the liver. These proteins serve to maintain the osmotic pressure and water balance as well as transport of materials 47. The present results are consistent with previous work that reported the change in serum protein in carcinogenic rats and attribute this to the impairment of the integrity of the cell membranes of the liver, kidney, and other tissues 3.
The present study shows that lycopene inhibited hepatic oxidative stress caused by DEN can ameliorate the biomarkers of liver injury and dysfunction in a dose-dependent manner (lycopene dose of 20 mg was the best treatment). Administration of lycopene (all groups) recorded significantly reduced transaminases, ALP, and TBIL in comparison with the DEN alone treated group. Lycopene also ameliorated the lowering effect of DEN on serum contents of TP and ALB. The present results agree with Bayramoglu et al. 6 who reported improvements in liver function parameters following treatment with lycopene in rats exposed to liver injury. The current finding together with available data suggest a protective impact of lycopene against liver cell damage induced by DEN, possibly due to its inhibitory effect on lipid peroxidation and its stimulatory effect on the antioxidant parameters, which led to stabilization of hepatocyte membrane and enhancement of liver synthetic function 48, 49. In addition, Cui et al. 9 reported that lipoproteins of lycopene can protect nucleic acids and proteins from oxidative stress that could lead to cancer. Moreover, the hepatoprotective effect of lycopene could be due to its ability to scavenge the free radicals and counteract its bad effects 10, 11.
The effect of DEN on lipid profile, as shown in the present work displayed significantly increased lipid fractions of TC, TG, and LDL-C, in contrast, to significantly reduced levels of HDL-C when compared to the control as previously reported 4, 50.
The effect of DEN on lipid profile, as shown in the present work displayed significantly increased lipid fractions of TC, TG, and LDL-C, in contrast, to significantly reduced levels of HDL-C when compared to the control as previously reported 4, 50. The present results indicated that lycopene exerted a lowering effect on DEN-induced hyperlipidemia in a dose-dependent manner. The obtained results displayed a significant decrease of TG, TC, and LDL-C; and a significant increase in HDL-C in serum of rats treated with lycopene especially the dose of 20 mg when compared to the group treated with DEN alone. The lipid-modulating effect of lycopene has previously been reported 51, 52. In addition, Inoue et al. 53 detected the ability of lycopene to significantly reduce serum TC and TG. Furthermore, the current finding of decreased level of LDL-C by lycopene is consistent with published data exhibiting a reduction in LDL-C levels following consumption of tomato products containing lycopene 54. Interestingly, the observed beneficial effect of lycopene on lipid metabolism raised its clinical efficacy as it could be used in the protection against cardiovascular diseases including atherosclerosis 11, 54. In this context, available data showed that lycopene may lower circulating cholesterol levels via increasing its fecal excretion and hence reducing its intestinal absorption. Also, it has been reported that lycopene may inhibit the synthesis of cholesterol and can protect LDL-C against oxidation thus preventing its contribution to the generation of atherosclerosis and may reduce the risk of cardiovascular diseases 51, 53.
The current data recorded significantly reduced all measured hematological parameters in all DEN treated groups. These findings are consistent with previous studies 4, 50 that attributed this reduction to the disturbance of the hematopoietic system and impairment of the cell membrane permeability which leading to osmotic swelling and cell hemolysis. Interestingly, the administration of lycopene to DEN treated groups attenuated the detrimental effects of DEN on hematological parameters in a dose-dependent manner that could be due to increased antioxidants that led to reduced oxidative stress on hematopoietic cells production or maturation systems 6, 55.
Regarding the immunohistochemical localization of the pro-inflammatory cytokines (TNF-α, IL-6) and α-fetoprotein tumor (AFP) marker protein expressions, the present study revealed upregulation of cytoplasmic reactivity of TNF-α, IL-6, and α-fetoprotein expressions in rat livers subjected to DEN. Cytokines play an important role in regulating immunity and include pro-inflammatory cytokines (TNF-α, IL-6, and IL-Iβ) 56. The inflammatory status due to DEN administration was confirmed in the present study by higher circulatory and tissue levels of TNF-α and IL-6 in the rats which hint that the inflammatory reaction appears after rats exposed to DEN as agreed with many previous studies 57, 58. The AFP was the most widely used tumor biomarker available for the early detection of HCC 59. It has long been recognized that the exposure of rats to certain carcinogens like DEN caused an increase of the circulating AFP levels associated with HCC 20. The upregulation of the AFP gene expression in DEN-administered rats might be due to the necrosis of hepatocytes that regulate the AFP synthesis on the cellular level 60. Although the mechanism of this regulation was not fully understood, the possible explanation for the initiation of AFP synthesis by neoplastic hepatocytes included either increased transcription of AFP gene or post-translational modification affecting AFP production 61.
Oxidative and inflammatory insults are intimately connected with each other in multistage carcinogenesis. Hence, it is expected that an agent with anti-inflammatory property will inhibit oxidative stress and vice versa 62. In agreement with the present study, Kim et al. 63 recorded the lycopene capability to scavenge free radicals in different systems and chelating metal ions, indicating that it can act as a natural antioxidant through many potential pathways and may be a useful therapeutic agent for treating radical-related pathological damage. Therefore, lycopene may be further developed as a potent antioxidant and anti-inflammatory agent in functional health food 64, 65. In agreement with the present AFP results, lycopene has been extensively studied for its antioxidant and cancer-preventing properties 66. In contrast to many other food phytonutrients whose effects have only been studied in animals, lycopene from tomatoes has been repeatedly studied in humans and found to be protective against several cancers, which now include colorectal, prostate, breast, lung, liver, and pancreatic cancers 67, 68 as agreeing with the present study.
Finally, the histopathological analysis of the liver tissue in the present study shows that the groups of animals treated with DEN recorded marked cancerous features which were ameliorated in the DEN treated groups supplemented with lycopene as consistent with a previous study 22.
The data of the present study proved that oral lycopene supplementation ameliorated all the investigated parameters in animals exposed to DEN. Moreover, lycopene was able to mitigate liver tissue damage induced by DEN through amelioration of the morphometric, histopathological, immunohistochemical, liver function, lipid profile, and hematological parameters. These findings suggested the potential efficacy of lycopene as an additional chemopreventive agent in the treatment of hepatocellular carcinoma by mitigating the apoptotic, oxidative stress and inflammatory effects as well as stimulation of carcinogen detoxification.
We are profoundly grateful to Prof. Dr. Sayed Abdel Raheem, Professor of Histopathology, Al- Azhar Faculty of Medicine, Cairo for his kind help during the histopathological investigation of colon tissues. A special thanks are given to Dr. Abdallah Hussien Fathy (Ph D), Laboratory Animal Facility Managing Director, Faculty of Pharmacy, Ain Shams University, Cairo 11566, Egypt for his great efforts in checking the plagiarism of our current manuscript.
The authors declare that they do not have any conflict of interest.
The authors declare that they do not have any source of funding.
| [1] | El-Serag, H. B. and Rudolph, K. L., Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology, 2007. 132(7): 2557-2576. | ||
| In article | View Article PubMed | ||
| [2] | Ferlay, J., Soerjomataram, I., Ervik, M., Dikshit, R., Eser, S., and Mathers, C., GLOBOCAN 2012: Cancer incidence and mortality, version 1.1. IARC Cancer Base ed. Lyon: IARC; 2014. | ||
| In article | |||
| [3] | Fathy, A. H., Bashandy, M. A., Bashandy, S. A. E., Mansour, A. M., and Elsadek, B., Sequential analysis and staging of a diethylnitrosamine induced hepatocellular carcinoma in male Wistar albino rat model. Can. J. Physiol. Pharmacol., 2017. 95(12): 1462-1472. | ||
| In article | View Article PubMed | ||
| [4] | Elsadek, B., Mansour, A., Saleem, T., Warnecke, A., and Kratz, F., The antitumor activity of a lactosaminated albumin conjugate of doxorubicin in a chemically induced hepatocellular carcinoma rat model compared to sorafenib. Dig. Liver Dis., 2017. 49(2): 213-222. | ||
| In article | View Article PubMed | ||
| [5] | Bansal, A. K., Bansal, M., Soni, G., and Bhatnagar, D., Modulation of N-nitrosodiethylamine (NDEA) induced oxidative stress by vitamin E in rat erythrocytes. Hum. Exp. Toxicol., 2005. 24(6): 297-302. | ||
| In article | View Article PubMed | ||
| [6] | Bayramoglu, G., Bayramoglu, A., Altuner, Y., Uyanoglu, M., and Colak, S., The effects of lycopene on hepatic ischemia/ reperfusion injury in rats. Cytotechnology, 2015. 67(3): 487-491. | ||
| In article | View Article PubMed | ||
| [7] | Hassan, H. F., Mansour, A. M., Abo‐Youssef, A. M., Elsadek, B. E., and Messiha, B. A., Zinc oxide nanoparticles as a novel anticancer approach; in vitro and in vivo evidence. Clin. Exp. Pharmacol. Physiol., 2017. 44(2): 235-243. | ||
| In article | View Article PubMed | ||
| [8] | Jamshidzadeh, A., Baghban, M., Azarpira, N., Bardbori, A. M., and Niknahad, H., Effects of tomato extract on oxidative stress induced toxicity in different organs of rats. Food Chem. Toxicol., 2008. 46(12): 3612-3615. | ||
| In article | View Article PubMed | ||
| [9] | Cui, L., Xu, F., Wu, K., Li, L., Qiao, T., Li, Z., Chen, T., and Sun, C., Anticancer effects and possible mechanisms of lycopene intervention on N-methylbenzylnitrosamine induced esophageal cancer in F344 rats based on PPARγ1. Eur. J. Pharmacol., 2020. 15(881): 173230. | ||
| In article | View Article PubMed | ||
| [10] | Abdel-Rahman, H. G., Abdelrazek, H., Zeidan, D. W., Mohamed, R. M., and Abdelazim, A. M., Lycopene: hepatoprotective and antioxidant effects toward bisphenol A-induced toxicity in female Wistar rats. Oxid. Med. Cell. Longev., 2018. 2018: Article ID 5167524, 8 pages. | ||
| In article | View Article PubMed | ||
| [11] | Fan, S., Sun, J. B., Li, R., Song, X., and Li, J., Lycopene protects myocardial ischemia injury through anti-apoptosis and anti-oxidative stress. Eur. Rev. Med. Pharmacol. Sci., 2019. 23(7): 3096-3104. | ||
| In article | |||
| [12] | Song, X., Luo, Y., Ma, L., Hu, X., Simal-Gandara, J., Wang, L. S., Bajpai, V. K., Xiao, J., and Chen, F., Recent trends and advances in the epidemiology, synergism, and delivery system of lycopene as an anti-cancer agent. In Seminars in Cancer Biology, 2021. Academic Press. | ||
| In article | View Article PubMed | ||
| [13] | Matos, H. R., Di Mascio, P., and Medeiros, M. H., Protective effect of lycopene on lipid peroxidation and oxidative DNA damage in cell culture. Arch. Biochem. Biophys., 2000. 383: 56-59. | ||
| In article | View Article PubMed | ||
| [14] | Rao, A. V. and Shen, H., Effect of low dose lycopene intake on lycopene bioavailability and oxidative stress. Nut. Res., 2002. 22(10): 1125-1131. | ||
| In article | View Article | ||
| [15] | Hsu, S.-M., Raine, L., and Fanger, H., Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. Journal of Histochemistry & Cytochemistry, 1981. 29(4): 577-580. | ||
| In article | View Article PubMed | ||
| [16] | Bancroft, J. D. and Gamble, M., editors. Theory and practice of histological techniques. Elsevier health sciences, 2008. | ||
| In article | |||
| [17] | Chakraborty, T., Chatterjee, A., Rana, A., Dhachinamoorthi, D., Kumar, A., and Chatterjee, M., Carcinogen-induced early molecular events and its implication in the initiation of chemical hepatocarcinogenesis in rats: Chemopreventive role of vanadium on this process. Biochim. Biophys. Acta. Mol. Basis Dis., 2007. 1772(1): 48-59. | ||
| In article | View Article PubMed | ||
| [18] | Hahn, W. C. and Weinberg, R. A., Rules for making human tumor cells. N. Engl. J. Med., 2002. 347(20): p. 1593-1603. | ||
| In article | View Article PubMed | ||
| [19] | Alison, M. R., Liver stem cells. Stem Cell Rev., 2005. 1(3): 253-260. | ||
| In article | View Article | ||
| [20] | Shahat, A. A., Alsaid, M. S., Kotob, S. E., and Ahmed, H. H., Significance of Rumex vesicarius as anticancer remedy against hepatocellular carcinoma: A proposal-based on experimental animal studies. Asian Pac. J. Cancer Prev., 2015. 16(10): 4303-4310. | ||
| In article | View Article PubMed | ||
| [21] | Sithithaworn, P., Yongvanit, P., Duenngai, K., Kiatsopit, N., and Pairojkul, C., Roles of liver fluke infection as risk factor for cholangiocarcinoma. J. Hepatobiliary Pancreat. Sci., 2014. 21(5): 301-308. | ||
| In article | View Article PubMed | ||
| [22] | Emre, K., Yilmaz, S., ÇERİBAŞI, A. O., and Selda, T., Protective effect of lycopene on diethylnitrosamine-induced oxidative stress and catalase expression in rats. Ankara Univ. Vet. Fak. Derg., 2018. 66(1): 43-52. | ||
| In article | View Article | ||
| [23] | Abdallah, I. Z. A. and Khattab, H. A. H., Protective role of lycopene against diethylnitrosamine induced experimental hepatocarcinogenesis. Egypt. J. Hosp. Med., 2004. 16(1): 1-13. | ||
| In article | View Article | ||
| [24] | Gupta, M., Mazumder, U. K., Kumar, R. S., Sivakumar, T., and Vamsi, M. L., Antitumor activity and antioxidant status of Caesalpinia bonducella against Ehrlich ascites carcinoma in Swiss albino mice. J. Pharmacol. Sci., 2004. 94(2): 177-184. | ||
| In article | View Article PubMed | ||
| [25] | Yilmaz, S., Atessahin, A., Sahna, E., Karahan, I., and Ozer, S., Protective effect of lycopene on adriamycin-induced cardiotoxicity and nephrotoxicity. Toxicology, 2006. 218(2-3): 164-171. | ||
| In article | View Article PubMed | ||
| [26] | Mayer, S. A. and Kulkarni, A. P., Hepatotoxicity. In: Hodgson E, Smart RC (eds) Introduction to biochemical toxicology. Wiley, New York, pp 599-628. 2001. | ||
| In article | |||
| [27] | Shaarawy, S. M., Tohamy, A. A., and Elgendy, S. M., Protective effects of garlic and silymarin on NDEA-induced rats hepato-toxicity. Int. J. Biol. Sci., 2009. 5: 549-557. | ||
| In article | View Article PubMed | ||
| [28] | Nordberg, J. and Arner, E. S., Reactive oxygen species, antioxidants, and the mammalian thioredoxin system. Free Radic. Biol. Med., 2001. 31(11): 1287-1312. | ||
| In article | View Article | ||
| [29] | Ramakrishnan, G., Raghavendran, H. R., and Vinodhkumar, R., Suppression of N-nitrosodiethylamine induced hepatocarcin-ogenesis by silymarin in rats. Chem. Biol. Interact., 2006. 161: 104-114. | ||
| In article | View Article PubMed | ||
| [30] | Dhanasekaran, M., Baskar, A. A., Ignacimuthu, S., Agastian, P., and Duraipandiyan, V., Chemopreventive potential of Epoxy clerodane diterpene from Tinospora cordifolia against diethylnitrosamine-induced hepatocellular carcinoma. Invest. New Drugs, 2009. 27(4): 347-355. | ||
| In article | View Article PubMed | ||
| [31] | Zahran, A. M., Azab, K. h., and Abbady, M. I., Modulatory role of allopurinol on xanthine oxidoreductase system and antioxidant status in irradiated rats. Egypt J. Rad. Sci. Appl., 2006. 19(2): 371-388. | ||
| In article | |||
| [32] | Seifried, H. E., McDonald, S. S., Anderson, D. E., Greenwald, P., and Milner, J. A., The antioxidant conundrum in cancer. Cancer Res., 2003. 63(15): 4295-4298. | ||
| In article | |||
| [33] | Vasquez-Garzon, V. R., Arellanes-Robledo, J., and Garcia-Roman, R., Inhibition of reactive oxygen species and pre-neoplastic le-sions by quercetin through an antioxidant defense mechanism. Free Radic. Res., 2009. 43: 128-137. | ||
| In article | View Article PubMed | ||
| [34] | Rajendra, P., Venugopal, M., Vasudev, V., and Pugalendi, K., Radioprotective effect of seasamol on γ-radiation induced DNA damage, lipid peroxidation and antioxidants level in cultured human lymphocytes. Toxicology, 2005. 209: 225-235. | ||
| In article | View Article PubMed | ||
| [35] | Zhao, B., Liu, H., Wang, J., Liu, P., Tan, X., Ren, B., Liu, Z., and Liu, X., Lycopene supplementation attenuates oxidative stress, neuroinflammation, and cognitive impairment in aged CD-1 mice. J. Agric. Food Chem., 2018. 66(12): 3127-3136. | ||
| In article | View Article PubMed | ||
| [36] | Altaş, S., Kızıl, G., Kızıl, M., Ketani, A., and Haris, P. I., Protective effect of Diyarbakır watermelon juice on carbon tetrachloride-induced toxicity in rats. Food Chem. Toxicol., 2011. 49(9): 2433-2438. | ||
| In article | View Article PubMed | ||
| [37] | Aydın, S., Tokaç, M., Taner, G., Arıkök, A. T., Dündar, H. Z., Özkardeş, A. B., Taşlıpınar, M. Y., Kılıç, M., Başaran, A. A., and Başaran, N., Antioxidant and antigenotoxic effects of lycopene in obstructive jaundice. J. Surg. Res., 2013. 182(2): 285-295. | ||
| In article | View Article PubMed | ||
| [38] | Lim, S., Hwang, S., Yu, J. H., Lim, J. W., and Kim, H., Lycopene inhibits regulator of calcineurin 1‐mediated apoptosis by reducing oxidative stress and down‐regulating Nucling in neuronal cells. Mol. Nutr. Food Res., 2017. 61(5): 1600530. | ||
| In article | View Article PubMed | ||
| [39] | Hashem, H. and Hussein, S. A., Lycopene mitigates experimental colitis in rats by inhibiting oxidative stress-mediated inflammation and apoptosis. Benha Vet. Med. J., 2020. 39(1): 16-21. | ||
| In article | View Article | ||
| [40] | Kurcer, M. A., Kurcer, Z., Koksal, M., Baba, F., Ocak, A. R., Aksoy, N., Atessahin, A., and Sahna, E., Effect of lycopene on caspase-3 enzyme activation in liver of methanol-intoxicated rats: comparison with fomepizole. J. Med. Food, 2010. 13(4): 985-991. | ||
| In article | View Article PubMed | ||
| [41] | Allard, W. J., Matera, J., Miller, M. C., Repollet, M., Connelly, M. C., Rao, C., Tibbe, A. G., Uhr, J. W., and Terstappen, L. W., Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin. Cancer Res., 2004. 10(20): 6897-6904. | ||
| In article | View Article PubMed | ||
| [42] | Tang, X., Yang, X., Peng, Y., and Lin, J., Protective effects of lycopene against H 2 O 2-induced oxidative injury and apoptosis in human endothelial cells. Cardiovasc. Drugs Ther., 2009. 23(6): 439-448. | ||
| In article | View Article PubMed | ||
| [43] | Gupta, V. K., Agarwal, S., Singh, P., and Pathania, D., Acrylic acid grafted cellulosic Luffa cylindrical fiber for the removal of dye and metal ions. Carbohydr. Polym., 2013. 98(1): 1214-1221. | ||
| In article | View Article PubMed | ||
| [44] | Elsayed, A., Elkomy, A., Elkammar, R., Youssef, G., Abdelhiee, E. Y., Abdo, W., Fadl, S. E., Soliman, A., and Aboubakr, M., Synergistic protective effects of lycopene and N-acetylcysteine against cisplatin-induced hepatorenal toxicity in rats. Sci. Rep., 2021. 11(1): 1-10. | ||
| In article | View Article PubMed | ||
| [45] | Bashandy, M. A., Abd El- Rasheid, H. G., Hasan, H. F., and Fathy, A. H., Protective and therapeutic effects of olive oil and Ficus carica as natural antioxidants on some biochemical parameters in liver of γ-irradiated male albino rats Al Azhar Bulletin of science, 2014. 25(1): 1-16. | ||
| In article | View Article | ||
| [46] | Carl, B. and David, B., Tietz fundamentals of clinical chemistry. Volume (1), fifth Edition, W.B. Saunders Company, USA. 2001. | ||
| In article | |||
| [47] | Imamoto, R., Okano, J. I., Sawada, S., Fujise, Y., Abe, R., and Murawaki, Y., Null anticarcinogenic effect of silymarin on diethylnitrosamine-induced hepatocarcinogenesis in rats. Exp. Ther. Med., 2014. 7(1): 31-38. | ||
| In article | View Article PubMed | ||
| [48] | Sahin, K., Orhan, C., Tuzcu, M., Sahin, N., Ali, S., Bahcecioglu, I. H., Guler, O., Ozercan, I., Ilhan, N., and Kucuk, O., Orally administered lycopene attenuates diethylnitrosamine-induced hepatocarcinogenesis in rats by modulating Nrf-2/HO-1 and Akt/mTOR pathways. Nutr. Cancer, 2014. 66(4): 590-598. | ||
| In article | View Article PubMed | ||
| [49] | Koul, A., Kaur, J., and Chugh, N. A., Protective Potential of Lycopene Enriched Tomato Extract against Dexamethasone Induced Hepatic and Renal Damage in Mice. Asian J. Biochem., 2021: 1-22. | ||
| In article | View Article | ||
| [50] | Fathy, A. H., Bashandy, M. A., Bashandy, S. A., Mansour, A. M., and Azab, K. S., The beneficial effect of natural antioxidants from olive oil with fig and date palm fruit extracts on biochemical and hematological parameters in rats treated with doxorubicin and γ-radiation. FACETS, 2018. 3(1):722-735. | ||
| In article | View Article | ||
| [51] | Elgawish, R. A., El-Beltagy, M. A., El-Sayed, R. M., Gaber, A. A., and Abdelrazek, H. M., Protective role of lycopene against metabolic disorders induced by chronic bisphenol A exposure in rats. Environ. Sci. Pollut. Res., 2020. 27(9): 9192-9201. | ||
| In article | View Article PubMed | ||
| [52] | Greish, S. M., Kader, G. S. A., Abdelaziz, E. Z., Eltamany, D. A., Sallam, H. S., and Abogresha, N. M., Lycopene is superior to moringa in improving fertility markers in diet-induced obesity male rats. Saudi J. Biol. Sci., 2021. 28(5): 2956-2963. | ||
| In article | View Article PubMed | ||
| [53] | Inoue, T., Yoshida, K., Sasaki, E., Aizawa, K., and Kamioka, H., Effects of lycopene intake on HDL-cholesterol and triglyceride levels: A systematic review with meta-analysis. J. Food Sci., 2021. 86(8):3285-3302. | ||
| In article | View Article PubMed | ||
| [54] | Liu, H., Liu, J., Liu, Z., Wang, Q., Liu, J., Feng, D., and Zou, J., Lycopene Reduces Cholesterol Absorption and Prevents Atherosclerosis in ApoE–/–Mice by Downregulating HNF-1α and NPC1L1 Expression. J. Agric. Food Chem., 2021. 69: 10114-10120. | ||
| In article | View Article PubMed | ||
| [55] | Hassan, H. A. and Edrees, G. M., Therapeutic effect of lycopene-rich tomato juice on cardiac disorder in rats fed on fried food in oxidized frying oil. Egypt. J. Hosp. Med., 2004. 14(1): 115-126. | ||
| In article | View Article | ||
| [56] | Libby, P., Ridker, P. M., and Maseri, A., Inflammation and atherosclerosis. Circulation, 2002. 105(9): 1135-1143. | ||
| In article | View Article PubMed | ||
| [57] | Abd-Elbaset, M., Mansour, A. M., Ahmed, O. M., and Abo-Youssef, A. M., The potential chemotherapeutic effect of β-ionone and/or sorafenib against hepatocellular carcinoma via its antioxidant effect, PPAR-γ, FOXO-1, Ki-67, Bax, and Bcl-2 signaling pathways. Naunyn-Schmiedeberg's Archives of Pharmacology, 2020. 393(9): 1611-1624. | ||
| In article | View Article PubMed | ||
| [58] | Bishayee, A., Barnes, K. F., Bhatia, D., Darvesh, A. S., and Carroll, R. T., Resveratrol suppresses oxidative stress and inflammatory response in diethylnitrosamine-initiated rat hepatocarcinogenesis. Cancer prevention research, 2010. 3(6): 753-763. | ||
| In article | View Article PubMed | ||
| [59] | El-Serag, H. B., Hepatocellular carcinoma. . N. Engl. J. Med., 2011. 365: 1118-1127. | ||
| In article | View Article PubMed | ||
| [60] | Siegel, R., Ma, J., Zou, Z., and Jemal , A., Cancer statistics. CA Cancer J. Clin., 2014. 64: 9-29. | ||
| In article | View Article PubMed | ||
| [61] | Song, P., Cai, Y., Tang, H., Li, C., and Huang, J., The clinical management of hepatocellular carcinoma worldwide: A concise review and comparison of current guidelines from 2001 to 2017. Biosci. Trends, 2017. 11(4): 389-398. | ||
| In article | View Article PubMed | ||
| [62] | Surh, Y.-J., NF-kappa B and Nrf2 as potential chemopreventive targets of some anti-inflammatory and antioxidative phytonutrients with anti-inflammatory and antioxidative activities. Asia Pac J Clin Nutr, 2008. 17(Suppl 1): 269-272. | ||
| In article | |||
| [63] | Kim, C. H., Park, M. K., Kim, S. K., and Cho, Y. H., Antioxidant capacity and anti‐inflammatory activity of lycopene in watermelon. International Journal of Food Science & Technology, 2014. 49(9): 2083-2091. | ||
| In article | View Article | ||
| [64] | Tricker, A. R. and Preussmann, R., Carcinogenic N-nitrosamines in the diet: occurrence, formation, mechanisms and carcinogenic potential. Mutat. Res., 1991. 259(3-4): 277-289. | ||
| In article | View Article | ||
| [65] | Cha, J. H., Kim, W. K., Ha, A. W., Kim, M. H., and Chang, M. J., Anti-inflammatory effect of lycopene in SW480 human colorectal cancer cells. Nutrition Research and Practice, 2017. 11(2): 90-96. | ||
| In article | View Article PubMed | ||
| [66] | Omoni, A. O. and Aluko, R. E., The anti-carcinogenic and anti-atherogenic effects of lycopene: a review. Trends in Food Science & Technology, 2005. 16(8): 344-350. | ||
| In article | View Article | ||
| [67] | Singh, P. and Goyal, G., Dietary lycopene: Its properties and anticarcinogenic effects. Comprehensive Reviews in Food Science and Food Safety, 2008. 7(3): 255-270. | ||
| In article | View Article PubMed | ||
| [68] | Friedman, M., Anticarcinogenic, cardioprotective, and other health benefits of tomato compounds lycopene, α-tomatine, and tomatidine in pure form and in fresh and processed tomatoes. Journal of agricultural and food chemistry, 2013. 61(40): 9534-9550. | ||
| In article | View Article PubMed | ||
| [1] | El-Serag, H. B. and Rudolph, K. L., Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology, 2007. 132(7): 2557-2576. | ||
| In article | View Article PubMed | ||
| [2] | Ferlay, J., Soerjomataram, I., Ervik, M., Dikshit, R., Eser, S., and Mathers, C., GLOBOCAN 2012: Cancer incidence and mortality, version 1.1. IARC Cancer Base ed. Lyon: IARC; 2014. | ||
| In article | |||
| [3] | Fathy, A. H., Bashandy, M. A., Bashandy, S. A. E., Mansour, A. M., and Elsadek, B., Sequential analysis and staging of a diethylnitrosamine induced hepatocellular carcinoma in male Wistar albino rat model. Can. J. Physiol. Pharmacol., 2017. 95(12): 1462-1472. | ||
| In article | View Article PubMed | ||
| [4] | Elsadek, B., Mansour, A., Saleem, T., Warnecke, A., and Kratz, F., The antitumor activity of a lactosaminated albumin conjugate of doxorubicin in a chemically induced hepatocellular carcinoma rat model compared to sorafenib. Dig. Liver Dis., 2017. 49(2): 213-222. | ||
| In article | View Article PubMed | ||
| [5] | Bansal, A. K., Bansal, M., Soni, G., and Bhatnagar, D., Modulation of N-nitrosodiethylamine (NDEA) induced oxidative stress by vitamin E in rat erythrocytes. Hum. Exp. Toxicol., 2005. 24(6): 297-302. | ||
| In article | View Article PubMed | ||
| [6] | Bayramoglu, G., Bayramoglu, A., Altuner, Y., Uyanoglu, M., and Colak, S., The effects of lycopene on hepatic ischemia/ reperfusion injury in rats. Cytotechnology, 2015. 67(3): 487-491. | ||
| In article | View Article PubMed | ||
| [7] | Hassan, H. F., Mansour, A. M., Abo‐Youssef, A. M., Elsadek, B. E., and Messiha, B. A., Zinc oxide nanoparticles as a novel anticancer approach; in vitro and in vivo evidence. Clin. Exp. Pharmacol. Physiol., 2017. 44(2): 235-243. | ||
| In article | View Article PubMed | ||
| [8] | Jamshidzadeh, A., Baghban, M., Azarpira, N., Bardbori, A. M., and Niknahad, H., Effects of tomato extract on oxidative stress induced toxicity in different organs of rats. Food Chem. Toxicol., 2008. 46(12): 3612-3615. | ||
| In article | View Article PubMed | ||
| [9] | Cui, L., Xu, F., Wu, K., Li, L., Qiao, T., Li, Z., Chen, T., and Sun, C., Anticancer effects and possible mechanisms of lycopene intervention on N-methylbenzylnitrosamine induced esophageal cancer in F344 rats based on PPARγ1. Eur. J. Pharmacol., 2020. 15(881): 173230. | ||
| In article | View Article PubMed | ||
| [10] | Abdel-Rahman, H. G., Abdelrazek, H., Zeidan, D. W., Mohamed, R. M., and Abdelazim, A. M., Lycopene: hepatoprotective and antioxidant effects toward bisphenol A-induced toxicity in female Wistar rats. Oxid. Med. Cell. Longev., 2018. 2018: Article ID 5167524, 8 pages. | ||
| In article | View Article PubMed | ||
| [11] | Fan, S., Sun, J. B., Li, R., Song, X., and Li, J., Lycopene protects myocardial ischemia injury through anti-apoptosis and anti-oxidative stress. Eur. Rev. Med. Pharmacol. Sci., 2019. 23(7): 3096-3104. | ||
| In article | |||
| [12] | Song, X., Luo, Y., Ma, L., Hu, X., Simal-Gandara, J., Wang, L. S., Bajpai, V. K., Xiao, J., and Chen, F., Recent trends and advances in the epidemiology, synergism, and delivery system of lycopene as an anti-cancer agent. In Seminars in Cancer Biology, 2021. Academic Press. | ||
| In article | View Article PubMed | ||
| [13] | Matos, H. R., Di Mascio, P., and Medeiros, M. H., Protective effect of lycopene on lipid peroxidation and oxidative DNA damage in cell culture. Arch. Biochem. Biophys., 2000. 383: 56-59. | ||
| In article | View Article PubMed | ||
| [14] | Rao, A. V. and Shen, H., Effect of low dose lycopene intake on lycopene bioavailability and oxidative stress. Nut. Res., 2002. 22(10): 1125-1131. | ||
| In article | View Article | ||
| [15] | Hsu, S.-M., Raine, L., and Fanger, H., Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. Journal of Histochemistry & Cytochemistry, 1981. 29(4): 577-580. | ||
| In article | View Article PubMed | ||
| [16] | Bancroft, J. D. and Gamble, M., editors. Theory and practice of histological techniques. Elsevier health sciences, 2008. | ||
| In article | |||
| [17] | Chakraborty, T., Chatterjee, A., Rana, A., Dhachinamoorthi, D., Kumar, A., and Chatterjee, M., Carcinogen-induced early molecular events and its implication in the initiation of chemical hepatocarcinogenesis in rats: Chemopreventive role of vanadium on this process. Biochim. Biophys. Acta. Mol. Basis Dis., 2007. 1772(1): 48-59. | ||
| In article | View Article PubMed | ||
| [18] | Hahn, W. C. and Weinberg, R. A., Rules for making human tumor cells. N. Engl. J. Med., 2002. 347(20): p. 1593-1603. | ||
| In article | View Article PubMed | ||
| [19] | Alison, M. R., Liver stem cells. Stem Cell Rev., 2005. 1(3): 253-260. | ||
| In article | View Article | ||
| [20] | Shahat, A. A., Alsaid, M. S., Kotob, S. E., and Ahmed, H. H., Significance of Rumex vesicarius as anticancer remedy against hepatocellular carcinoma: A proposal-based on experimental animal studies. Asian Pac. J. Cancer Prev., 2015. 16(10): 4303-4310. | ||
| In article | View Article PubMed | ||
| [21] | Sithithaworn, P., Yongvanit, P., Duenngai, K., Kiatsopit, N., and Pairojkul, C., Roles of liver fluke infection as risk factor for cholangiocarcinoma. J. Hepatobiliary Pancreat. Sci., 2014. 21(5): 301-308. | ||
| In article | View Article PubMed | ||
| [22] | Emre, K., Yilmaz, S., ÇERİBAŞI, A. O., and Selda, T., Protective effect of lycopene on diethylnitrosamine-induced oxidative stress and catalase expression in rats. Ankara Univ. Vet. Fak. Derg., 2018. 66(1): 43-52. | ||
| In article | View Article | ||
| [23] | Abdallah, I. Z. A. and Khattab, H. A. H., Protective role of lycopene against diethylnitrosamine induced experimental hepatocarcinogenesis. Egypt. J. Hosp. Med., 2004. 16(1): 1-13. | ||
| In article | View Article | ||
| [24] | Gupta, M., Mazumder, U. K., Kumar, R. S., Sivakumar, T., and Vamsi, M. L., Antitumor activity and antioxidant status of Caesalpinia bonducella against Ehrlich ascites carcinoma in Swiss albino mice. J. Pharmacol. Sci., 2004. 94(2): 177-184. | ||
| In article | View Article PubMed | ||
| [25] | Yilmaz, S., Atessahin, A., Sahna, E., Karahan, I., and Ozer, S., Protective effect of lycopene on adriamycin-induced cardiotoxicity and nephrotoxicity. Toxicology, 2006. 218(2-3): 164-171. | ||
| In article | View Article PubMed | ||
| [26] | Mayer, S. A. and Kulkarni, A. P., Hepatotoxicity. In: Hodgson E, Smart RC (eds) Introduction to biochemical toxicology. Wiley, New York, pp 599-628. 2001. | ||
| In article | |||
| [27] | Shaarawy, S. M., Tohamy, A. A., and Elgendy, S. M., Protective effects of garlic and silymarin on NDEA-induced rats hepato-toxicity. Int. J. Biol. Sci., 2009. 5: 549-557. | ||
| In article | View Article PubMed | ||
| [28] | Nordberg, J. and Arner, E. S., Reactive oxygen species, antioxidants, and the mammalian thioredoxin system. Free Radic. Biol. Med., 2001. 31(11): 1287-1312. | ||
| In article | View Article | ||
| [29] | Ramakrishnan, G., Raghavendran, H. R., and Vinodhkumar, R., Suppression of N-nitrosodiethylamine induced hepatocarcin-ogenesis by silymarin in rats. Chem. Biol. Interact., 2006. 161: 104-114. | ||
| In article | View Article PubMed | ||
| [30] | Dhanasekaran, M., Baskar, A. A., Ignacimuthu, S., Agastian, P., and Duraipandiyan, V., Chemopreventive potential of Epoxy clerodane diterpene from Tinospora cordifolia against diethylnitrosamine-induced hepatocellular carcinoma. Invest. New Drugs, 2009. 27(4): 347-355. | ||
| In article | View Article PubMed | ||
| [31] | Zahran, A. M., Azab, K. h., and Abbady, M. I., Modulatory role of allopurinol on xanthine oxidoreductase system and antioxidant status in irradiated rats. Egypt J. Rad. Sci. Appl., 2006. 19(2): 371-388. | ||
| In article | |||
| [32] | Seifried, H. E., McDonald, S. S., Anderson, D. E., Greenwald, P., and Milner, J. A., The antioxidant conundrum in cancer. Cancer Res., 2003. 63(15): 4295-4298. | ||
| In article | |||
| [33] | Vasquez-Garzon, V. R., Arellanes-Robledo, J., and Garcia-Roman, R., Inhibition of reactive oxygen species and pre-neoplastic le-sions by quercetin through an antioxidant defense mechanism. Free Radic. Res., 2009. 43: 128-137. | ||
| In article | View Article PubMed | ||
| [34] | Rajendra, P., Venugopal, M., Vasudev, V., and Pugalendi, K., Radioprotective effect of seasamol on γ-radiation induced DNA damage, lipid peroxidation and antioxidants level in cultured human lymphocytes. Toxicology, 2005. 209: 225-235. | ||
| In article | View Article PubMed | ||
| [35] | Zhao, B., Liu, H., Wang, J., Liu, P., Tan, X., Ren, B., Liu, Z., and Liu, X., Lycopene supplementation attenuates oxidative stress, neuroinflammation, and cognitive impairment in aged CD-1 mice. J. Agric. Food Chem., 2018. 66(12): 3127-3136. | ||
| In article | View Article PubMed | ||
| [36] | Altaş, S., Kızıl, G., Kızıl, M., Ketani, A., and Haris, P. I., Protective effect of Diyarbakır watermelon juice on carbon tetrachloride-induced toxicity in rats. Food Chem. Toxicol., 2011. 49(9): 2433-2438. | ||
| In article | View Article PubMed | ||
| [37] | Aydın, S., Tokaç, M., Taner, G., Arıkök, A. T., Dündar, H. Z., Özkardeş, A. B., Taşlıpınar, M. Y., Kılıç, M., Başaran, A. A., and Başaran, N., Antioxidant and antigenotoxic effects of lycopene in obstructive jaundice. J. Surg. Res., 2013. 182(2): 285-295. | ||
| In article | View Article PubMed | ||
| [38] | Lim, S., Hwang, S., Yu, J. H., Lim, J. W., and Kim, H., Lycopene inhibits regulator of calcineurin 1‐mediated apoptosis by reducing oxidative stress and down‐regulating Nucling in neuronal cells. Mol. Nutr. Food Res., 2017. 61(5): 1600530. | ||
| In article | View Article PubMed | ||
| [39] | Hashem, H. and Hussein, S. A., Lycopene mitigates experimental colitis in rats by inhibiting oxidative stress-mediated inflammation and apoptosis. Benha Vet. Med. J., 2020. 39(1): 16-21. | ||
| In article | View Article | ||
| [40] | Kurcer, M. A., Kurcer, Z., Koksal, M., Baba, F., Ocak, A. R., Aksoy, N., Atessahin, A., and Sahna, E., Effect of lycopene on caspase-3 enzyme activation in liver of methanol-intoxicated rats: comparison with fomepizole. J. Med. Food, 2010. 13(4): 985-991. | ||
| In article | View Article PubMed | ||
| [41] | Allard, W. J., Matera, J., Miller, M. C., Repollet, M., Connelly, M. C., Rao, C., Tibbe, A. G., Uhr, J. W., and Terstappen, L. W., Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin. Cancer Res., 2004. 10(20): 6897-6904. | ||
| In article | View Article PubMed | ||
| [42] | Tang, X., Yang, X., Peng, Y., and Lin, J., Protective effects of lycopene against H 2 O 2-induced oxidative injury and apoptosis in human endothelial cells. Cardiovasc. Drugs Ther., 2009. 23(6): 439-448. | ||
| In article | View Article PubMed | ||
| [43] | Gupta, V. K., Agarwal, S., Singh, P., and Pathania, D., Acrylic acid grafted cellulosic Luffa cylindrical fiber for the removal of dye and metal ions. Carbohydr. Polym., 2013. 98(1): 1214-1221. | ||
| In article | View Article PubMed | ||
| [44] | Elsayed, A., Elkomy, A., Elkammar, R., Youssef, G., Abdelhiee, E. Y., Abdo, W., Fadl, S. E., Soliman, A., and Aboubakr, M., Synergistic protective effects of lycopene and N-acetylcysteine against cisplatin-induced hepatorenal toxicity in rats. Sci. Rep., 2021. 11(1): 1-10. | ||
| In article | View Article PubMed | ||
| [45] | Bashandy, M. A., Abd El- Rasheid, H. G., Hasan, H. F., and Fathy, A. H., Protective and therapeutic effects of olive oil and Ficus carica as natural antioxidants on some biochemical parameters in liver of γ-irradiated male albino rats Al Azhar Bulletin of science, 2014. 25(1): 1-16. | ||
| In article | View Article | ||
| [46] | Carl, B. and David, B., Tietz fundamentals of clinical chemistry. Volume (1), fifth Edition, W.B. Saunders Company, USA. 2001. | ||
| In article | |||
| [47] | Imamoto, R., Okano, J. I., Sawada, S., Fujise, Y., Abe, R., and Murawaki, Y., Null anticarcinogenic effect of silymarin on diethylnitrosamine-induced hepatocarcinogenesis in rats. Exp. Ther. Med., 2014. 7(1): 31-38. | ||
| In article | View Article PubMed | ||
| [48] | Sahin, K., Orhan, C., Tuzcu, M., Sahin, N., Ali, S., Bahcecioglu, I. H., Guler, O., Ozercan, I., Ilhan, N., and Kucuk, O., Orally administered lycopene attenuates diethylnitrosamine-induced hepatocarcinogenesis in rats by modulating Nrf-2/HO-1 and Akt/mTOR pathways. Nutr. Cancer, 2014. 66(4): 590-598. | ||
| In article | View Article PubMed | ||
| [49] | Koul, A., Kaur, J., and Chugh, N. A., Protective Potential of Lycopene Enriched Tomato Extract against Dexamethasone Induced Hepatic and Renal Damage in Mice. Asian J. Biochem., 2021: 1-22. | ||
| In article | View Article | ||
| [50] | Fathy, A. H., Bashandy, M. A., Bashandy, S. A., Mansour, A. M., and Azab, K. S., The beneficial effect of natural antioxidants from olive oil with fig and date palm fruit extracts on biochemical and hematological parameters in rats treated with doxorubicin and γ-radiation. FACETS, 2018. 3(1):722-735. | ||
| In article | View Article | ||
| [51] | Elgawish, R. A., El-Beltagy, M. A., El-Sayed, R. M., Gaber, A. A., and Abdelrazek, H. M., Protective role of lycopene against metabolic disorders induced by chronic bisphenol A exposure in rats. Environ. Sci. Pollut. Res., 2020. 27(9): 9192-9201. | ||
| In article | View Article PubMed | ||
| [52] | Greish, S. M., Kader, G. S. A., Abdelaziz, E. Z., Eltamany, D. A., Sallam, H. S., and Abogresha, N. M., Lycopene is superior to moringa in improving fertility markers in diet-induced obesity male rats. Saudi J. Biol. Sci., 2021. 28(5): 2956-2963. | ||
| In article | View Article PubMed | ||
| [53] | Inoue, T., Yoshida, K., Sasaki, E., Aizawa, K., and Kamioka, H., Effects of lycopene intake on HDL-cholesterol and triglyceride levels: A systematic review with meta-analysis. J. Food Sci., 2021. 86(8):3285-3302. | ||
| In article | View Article PubMed | ||
| [54] | Liu, H., Liu, J., Liu, Z., Wang, Q., Liu, J., Feng, D., and Zou, J., Lycopene Reduces Cholesterol Absorption and Prevents Atherosclerosis in ApoE–/–Mice by Downregulating HNF-1α and NPC1L1 Expression. J. Agric. Food Chem., 2021. 69: 10114-10120. | ||
| In article | View Article PubMed | ||
| [55] | Hassan, H. A. and Edrees, G. M., Therapeutic effect of lycopene-rich tomato juice on cardiac disorder in rats fed on fried food in oxidized frying oil. Egypt. J. Hosp. Med., 2004. 14(1): 115-126. | ||
| In article | View Article | ||
| [56] | Libby, P., Ridker, P. M., and Maseri, A., Inflammation and atherosclerosis. Circulation, 2002. 105(9): 1135-1143. | ||
| In article | View Article PubMed | ||
| [57] | Abd-Elbaset, M., Mansour, A. M., Ahmed, O. M., and Abo-Youssef, A. M., The potential chemotherapeutic effect of β-ionone and/or sorafenib against hepatocellular carcinoma via its antioxidant effect, PPAR-γ, FOXO-1, Ki-67, Bax, and Bcl-2 signaling pathways. Naunyn-Schmiedeberg's Archives of Pharmacology, 2020. 393(9): 1611-1624. | ||
| In article | View Article PubMed | ||
| [58] | Bishayee, A., Barnes, K. F., Bhatia, D., Darvesh, A. S., and Carroll, R. T., Resveratrol suppresses oxidative stress and inflammatory response in diethylnitrosamine-initiated rat hepatocarcinogenesis. Cancer prevention research, 2010. 3(6): 753-763. | ||
| In article | View Article PubMed | ||
| [59] | El-Serag, H. B., Hepatocellular carcinoma. . N. Engl. J. Med., 2011. 365: 1118-1127. | ||
| In article | View Article PubMed | ||
| [60] | Siegel, R., Ma, J., Zou, Z., and Jemal , A., Cancer statistics. CA Cancer J. Clin., 2014. 64: 9-29. | ||
| In article | View Article PubMed | ||
| [61] | Song, P., Cai, Y., Tang, H., Li, C., and Huang, J., The clinical management of hepatocellular carcinoma worldwide: A concise review and comparison of current guidelines from 2001 to 2017. Biosci. Trends, 2017. 11(4): 389-398. | ||
| In article | View Article PubMed | ||
| [62] | Surh, Y.-J., NF-kappa B and Nrf2 as potential chemopreventive targets of some anti-inflammatory and antioxidative phytonutrients with anti-inflammatory and antioxidative activities. Asia Pac J Clin Nutr, 2008. 17(Suppl 1): 269-272. | ||
| In article | |||
| [63] | Kim, C. H., Park, M. K., Kim, S. K., and Cho, Y. H., Antioxidant capacity and anti‐inflammatory activity of lycopene in watermelon. International Journal of Food Science & Technology, 2014. 49(9): 2083-2091. | ||
| In article | View Article | ||
| [64] | Tricker, A. R. and Preussmann, R., Carcinogenic N-nitrosamines in the diet: occurrence, formation, mechanisms and carcinogenic potential. Mutat. Res., 1991. 259(3-4): 277-289. | ||
| In article | View Article | ||
| [65] | Cha, J. H., Kim, W. K., Ha, A. W., Kim, M. H., and Chang, M. J., Anti-inflammatory effect of lycopene in SW480 human colorectal cancer cells. Nutrition Research and Practice, 2017. 11(2): 90-96. | ||
| In article | View Article PubMed | ||
| [66] | Omoni, A. O. and Aluko, R. E., The anti-carcinogenic and anti-atherogenic effects of lycopene: a review. Trends in Food Science & Technology, 2005. 16(8): 344-350. | ||
| In article | View Article | ||
| [67] | Singh, P. and Goyal, G., Dietary lycopene: Its properties and anticarcinogenic effects. Comprehensive Reviews in Food Science and Food Safety, 2008. 7(3): 255-270. | ||
| In article | View Article PubMed | ||
| [68] | Friedman, M., Anticarcinogenic, cardioprotective, and other health benefits of tomato compounds lycopene, α-tomatine, and tomatidine in pure form and in fresh and processed tomatoes. Journal of agricultural and food chemistry, 2013. 61(40): 9534-9550. | ||
| In article | View Article PubMed | ||
