Anti-fibrotic Effects and Mechanism of Shengmai Injection （生脉注射液） on Human Hepatic Stellate Cells LX-2

• 来源期刊: 中国结合医学杂志（英文版）
• 年，卷（期）: 2019,（3）
• 页码范围: 197-202
• 页数: 6页
• 语种: 英文
• DOI: 10.1007/s11655-018-2849-x

摘要

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 × 104 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 6μ L/mL).Cell proliferation was measured after an additional 24 or 48 h using the 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (M T T) 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％ confluence,apoptosis was detected by flow 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 μ L/mL of SMI significantly inhibited cell proliferation at 24 and 48 h after treatment (P＜0.05).And 2.4μ L/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 μ L/mL (P＜0.05).Conclusions: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 6μ L/mL) could accelerate apoptosis in LX-2 cells,and the mechanism might be associated with NDRG2 over-expression.