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目的观察复方制剂丹芍化纤胶囊治疗大鼠CCl4中毒性肝纤维化对HSC细胞凋亡及细胞周期的影响,以探讨丹芍化纤胶囊可能的作用机理。方法雄性Wistar大鼠采用CCl4、饮酒、高脂低蛋白饮食等复合病因刺激制备肝纤维化动物模型8周,然后给予丹芍化纤胶囊(1g/kg)灌胃治疗8周。实验结束时测定肝脏指数、血清透明质酸(HA)及谷丙转氨酶(ALT)含量,光镜下观察肝组织纤维化程度,检测尿羟脯氨酸(Hyp)排出量,同时用流式细胞仪检测治疗前后肝星状细胞(HSC)凋亡并分析细胞周期各期细胞比例。结果治疗组大鼠的肝脏指数、血清HA、ALT含量明显低于肝纤维化模型组;肝纤维化程度明显轻于肝纤维化模型组;尿Hyp排出量显著高于肝纤维化模型组及自然恢复组;HSC细胞凋亡、G0-G1期细胞比例明显高于肝纤维化模型组(分别为7.81±0.47/0.28±0.05,94.30±1.33/68.40±6.47,P<0.05),S期细胞比例显著低于肝纤维化模型组及自然恢复组(分别为3.11±1.27,9.83±1.81,11.87±1.92,P<0.05)。结论丹芍化纤胶囊对大鼠CCl4中毒性肝纤维化有一定的治疗作用,这种治疗作用可能部分是通过抑制HSC细胞增殖及促进HSC细胞凋亡而致。
Objective To observe the effect of Danshaohuaxian Capsule, a compound preparation, on the apoptosis and cell cycle of HSC in rats with CCl4 toxic hepatic fibrosis, in order to investigate the possible mechanism of Danshaohuaxian capsule. METHODS Male Wistar rats were treated with compound etiology such as CCl4, drinking and high-fat low-protein diet to prepare an animal model of hepatic fibrosis for 8 weeks. Then, Danshaohuaxian capsules (1g/kg) were intragastrically administered for 8 weeks. At the end of the experiment, the liver index, serum HA, and alanine aminotransferase (ALT) levels were measured. The degree of hepatic fibrosis was observed under light microscopy, and the urinary hydroxyproline (Hyp) output was measured, while flow cytometry was used. Apoptosis of hepatic stellate cells (HSCs) was measured before and after treatment and the proportion of cells in each phase of the cell cycle was analyzed. Results The liver index, serum HA and ALT levels in the treatment group were significantly lower than those in the hepatic fibrosis model group. The liver fibrosis was significantly lighter than that in the hepatic fibrosis model group. The urinary Hyp excretion was significantly higher than the hepatic fibrosis model group and natural Recovery group; HSC cell apoptosis, G0-G1 phase cell ratio was significantly higher than the liver fibrosis model group (respectively 7.81 ± 0.47/0.28 ± 0.05, 94.30 ± 1.33/68.40 ± 6.47, P <0.05), S phase cell ratio Significantly lower than liver fibrosis model group and natural recovery group (3.11±1.27, 9.83±1.81, 11.87±1.92, respectively, P<0.05). Conclusion Danshaohuaxian capsule has a certain therapeutic effect on CCl4 toxic hepatic fibrosis in rats, and this therapeutic effect may be partly caused by inhibiting the proliferation of HSC cells and promoting the apoptosis of HSC cells.