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目的探讨黄芪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。黄芪注射液(0mg/ml、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。黄芪注射液(200mg/ml)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黄芪注射液对活化的肝星状细胞凋亡的影响。结果黄芪注射液各浓度组(除25mg/ml作用24h组外)OD值较对照组明显降低(P<0.05);黄芪注射液200,mg/ml组作用24和48h流式细胞术检测出G2-M期的细胞数均明显高于正常对照组(P<0.01);黄芪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。结论黄芪注射液主要是通过抑制肝星状细胞增殖的G2-M期而产生抗肝纤维化作用的。
Objective To investigate the anti-hepatic fibrosis effect of Huangqi injection and its mechanism. Methods The separation of hepatic stellate cells was performed by collagenase cycling perfusion. After treatment with Huangqi Injection (0mg/ml, 25mg/ml, 50mg/ml, 100mg/ml, 200mg/ml, 400mg/ml) for different time (24h, 48h, 72h), MTT assay was used to detect cell activation and proliferation. . After 24 h and 48 h of Astragalus injection (200 mg/ml), the cell proliferation cycle was detected by flow cytometry. The effect of astragalus injection on the apoptosis of activated hepatic stellate cells was detected by ethidium bromide/acridine orange fluorescence staining combined with microscopic observation and flow cytometry. Results Compared with the control group, the OD value of each concentration group of Huangqi injection (except 25mg/ml for 24h group) was significantly lower (P<0.05); the G2 was detected by flow cytometry 24h and 48h of Huangqi injection 200mg/ml group. The number of cells in the -M phase was significantly higher than that in the normal control group (P<0.01); the apoptosis of hepatic stellate cells was not detected by fluorescence staining and flow cytometry at each concentration of Huangqi injection. Conclusion Astragalus injection can inhibit liver fibrosis by inhibiting the G2-M phase of hepatic stellate cell proliferation.