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Curcumin Suppresses the Activity of Inhibitor-κB Kinase in an in vitro Inflamed Human Intestinal Mucosa Model by S-nitrosylation

Ning-Jo Kao, Zwe-Ling Kong

Journal of Food and Nutrition Research. 2017, 5(6), 406-412 doi:10.12691/jfnr-5-6-7
  • Figure 1. The cytotoxicity of curcumin on Caco-2 and HT-29 cells. Both Caoco-2 (A) and HT-29 cells (B) were incubated with samples for 12 h or 48 h and viability was assessed using an MTT assay. Experiments were repeated 3 times independently to ensure reproducibility and the standard deviation of the mean are represented as error bars (n=3)
  • Figure 2. The repressionof iNOS in LPS-induced human intestinal cells by curcumin. Total lysates was extracted from LPS-induced Caco-2 and HT-29 cellsof each experimental group at 12hindividually. The expression of iNOS was detected by Western blot, and β-actin was as the loading control. Experiments were repeated 3 times independently to ensure reproducibility and the standard deviation of the mean are represented as error bars (n=3). Values with different letters were significantly different (p< 0.05) at corresponding concentrations between different treatments
  • Figure 3. Inhibition of nitrite and nitrate in LPS-induced Caco-2 and HT-29 cells by curcumin. Nitrite and nitrate were detected from Caco-2 and HT-29cells cultured medium of each experimental group at 12hindividuallyby used Griess reagent(A-B).The data was repeated 3 times independently to ensure reproducibility and the standard deviation of the mean are represented as error bars (n=3). Values with different letters were significantly different (p< 0.05) at corresponding concentrations between different treatments
  • Figure 4. Cytokine expression in LPS-induced Caco-2 and HT-29 cells by curcumin. IL-1β, IL-6, TNF-αand IFN-γwere detected from Caco-2 and HT-29 cells cultured medium of each experimental group at 12hindividually (4A-B) by used microarray. The data was repeated 3 times independently to ensure reproducibility and the standard deviation of the mean are represented as error bars (n=3). Values with different letters were significantly different (p< 0.05) at corresponding concentrations between different treatments
  • Figure 5. Total S-nitrosylated protein expression in LPS-induced Caco-2 and HT-29 cells by treated with curcumin. The total S-nitrosylated protein extracted from Caco-2 and HT-29 cell lysates of each experimental group at12h individually that was prepared by biotin switch method and detected by Western blot using anti-biotin. The bar-chart presented the relative quantification of total S-nitrosylation from Western blot. The data was repeated 3 times independently to ensure reproducibility and the standard deviation of the mean are represented as error bars (n=3). Values with different letters were significantly different (p< 0.05) at corresponding concentrations between different treatments
  • Figure 6. S-nitrosylated IKK-β Expression in LPS-induced Caco-2 and HT-29 cells by treated with curcumin. S-nitrosylated IKKβ as obtained from cell lysates of each experimental group at12hindividually by biotin switch method and detected by Western blot using anti-biotin. The expression level of IKKβ was as the control and relative fold change of S-nitrosylated IKKβ was normalized by IKKβ protein expression. The data was repeated 3 times independently to ensure reproducibility and the standard deviation of the mean are represented as error bars (n=3). Values with different letters were significantly different (p< 0.05) at corresponding concentrations between different treatments
  • Figure7. IκBphosphorylation and NF-κB activation in LPS-induced Caco-2 and HT-29 cells by treated with curcumin. The expression level of phosphorylated IκB (pIκB), IκB, and cytosolic/nuclear NF-κB proteins were detected by Western blot. Actin expression was as the loading control. The bar chart presented the relative quantification of p-IκB expression normalized by IκB protein expression The data was repeated 3 times independently to ensure reproducibility and the standard deviation of the mean are represented as error bars (n=3). Values with different letters were significantly different (p< 0.05) at corresponding concentrations between different treatments