1. Anti-fibrotic Effects and Mechanism of Shengmai Injection (生脉注射液) on Human Hepatic Stellate Cells LX-2.
- Author
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Zhang, Yi, Ma, Li-tian, Li, Jie, Qiao, Yu, Liu, Jun-ye, Wang, Jin, Ren, Qin-you, Hu, Jin-tao, and Zheng, Jin
- Subjects
CELL proliferation ,APOPTOSIS ,BIOLOGICAL assay ,BROMIDES ,BUFFER solutions ,CELL lines ,CULTURE media (Biology) ,FLOW cytometry ,GENE expression ,HERBAL medicine ,LIVER ,CHINESE medicine ,MICROSCOPY ,ONCOGENES ,THIAZOLES ,FIBROSIS ,IN vitro studies ,DRUG administration ,DRUG dosage - Abstract
Objective: To investigate the effects of Shengmai Injection (生脉注射液, SMI) on the proliferation, apoptosis and N-myc downstream-regulated gene 2 (NDRG2, a tumour suppressor gene) expression in varying densities of human hepatic stellate cells LX-2.Methods: LX-2 cells were cultured in vitro. Then, cells were plated in 96-well plates at an approximate density of 2.5×10
4 cells/mL and cultured for 48, 72, 96 or 120 h followed by the application of different concentrations of SMI (0.6, 1.2, 2.4, 4.8 or 6 μL/mL). Cell proliferation was measured after an additional 24 or 48 h using the 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of SMI on different cell growth states (cultured for 48, 72, 96, or 120 h) were observed by light microscopy at 24 h after treatment. When the cells reached 80% conflfluence, apoptosis was detected by flflow cytometry after 24 h. Lastly, LX-2 cells were treated with different concentrations of SMI and extracted with protein lysis buffer. The levels of NDRG2 were measured by Western blot.Results: When the LX-2 cells grew for 48, 72, 96 and 120 h, 4.8 and 6 μL/mL of SMI significantly inhibited cell proliferation at 24 and 48 h after treatment (P<0.05). And 2.4 μL/mL of SMI also inhibited cell proliferation at 24 h after treatment when cell growth for 48 h (P<0.05) and at 48 h after treatment when cell growth for 72, 96 and 120 h (P<0.05). The NDRG2 expression level in the LX-2 cell was significantly increased when treated with SMI at concentrations of 1.2, 2.4, 4.8 or 6 μL/mL (P<0.05).Conclusion: The inhibitory effects of SMI on the proliferation of LX-2 cells were related to not only concentration dependent but also cell density. In addition, SMI (2.4, 4.8 and 6 μL/mL) could accelerate apoptosis in LX-2 cells, and the mechanism might be associated with NDRG2 over-expression. [ABSTRACT FROM AUTHOR]- Published
- 2019
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