硫普罗宁注射液干预大鼠肝星状细胞致肝纤维化形成作用的研究

目的 探讨硫普罗宁注射液抗肝纤维化的作用及其机制。方法 肝星状细胞的分离采用胶原酶循环灌流法。硫普罗宁注射液（25mg／ml、50mg／ml、100mg／ml、200mg／ml、400mg／ml）作用不同时间（24h、48h、72h）后，采用MTT法OD值检测细胞活化增殖情况。硫普罗宁注射液（200mg／m1）作用24h和48h后，流式细胞术检测细胞增殖周期。采用溴乙锭／吖啶橙荧光染色法结合显微镜下形态学观察及流式细胞术检测硫普罗宁注射液对活化的肝星状细胞凋亡的影响。结果 硫普罗宁注射液各浓度组OD值较对照组明显降低（P〈0.05），硫普罗宁注射液200mg／ml组作用24h和48h流式细胞术检测出C2-M期的细胞数均明显高于正常对照组（P〈0．01），硫普罗宁注射液各浓度组荧光染色法和流式细胞术均未检测到肝星状细胞凋亡。结论 硫普罗宁注射液主要是通过抑制肝星状细胞增殖的C2-M期而产生抗肝纤维化作用的。

The effects of tiopronin injection preventing hepatic fibrosis through inhibiting proliferation of cultured rat hepatic stellate cells

Objective To investigate the effects of Tiopronin injection on the proliferation and apoptpsis of hepatic stellate cells(HSC) in vitro.Methods HSCs were incubated with various concentration of Tiopronin injection(25mg/ml,50mg/ml,100mg/ml,200mg/ml,400mg/ml) for 24,48 and 72 hours.Cell proliferation was detected with MTT colorimetric assay.Cell proliferation cycle was detected with flow cytometry.Cell apoptosis was detected with AO/EB fluorescent staining and flow cytometry.Results OD values in every Tiopronin injection group(except for the 25mg/ml,24hour group) was significantly lower than that in control group(P<0.05).The cell numbers of G2-M phase detected by flow cytometry in Tiopronin injection 200mg/ml 24h and 48h group were significantly higher than that in control group(P<0.01).The apoptosis of HSCs was not detected in every Tiopronin ingection group through AO/EB fluorescent staining and flow cytometry.Conclusion Tiopronin injection can prevent hepatic fibrosis mainly through inhibiting proliferation of HSCs.