Figure 1. Dosage optimization of TGF-β1 to induce hypertrophic responses in primary neonatal rat cardiomyocytes (NRCMs). NRCMs were seeded at a density of 106 cells/well in 24-well plates and cultured overnight. After a 24- h incubation in serum-free medium, NRCMs were stimulated with 0, 0.1, 1, 3, or 10 μg/L of TGF-β1 for 24 h. (A–F) Cellular RNA was stained with RNA-sensitive fluorescence probe propidium iodide (PI) after DNase treatment. Representative images are shown in (A–E). Scale bar: 100 μm. The fluorescent signals were measured in five randomly selected fields in each well (F). (G) Protein synthesis was evaluated by measuring the incorporation of [3H]-leucine. The radioactivity of [3H]-leucine incorporated into the trichloroacetic acid precipitable material was determined by β-scintillation counting. The mRNA levels of atrial natriuretic factor (ANF) (H) and β-myosin heavy chain (β-MHC) (I) were determined using quantitative real-time PCR (qRT-PCR). GAPDH was used as an internal control. Data are expressed as mean ± standard deviation (SD). *P < 0.05 vs. control; #P < 0.05 vs. 3 μg/L TGF-β1 (n=7).


Puerarin Attenuates Transforming Growth Factor Beta-1 Induced Hypertrophic Responses and Smad Proetin Upregulation in Neonatal Rat Cardiomyocytes

Yongying Shi, Aijun Liu, Junjiang Lu, Guangyuan Chen, Shiming Liu, Minsheng Chen, Chengfeng Luo

American Journal of Biomedical Research. 2020, 8(2), 40-46 doi:10.12691/ajbr-8-2-3