三甲益肝颗粒含药血清对HSC-T6细胞形态,增殖和凋亡的影响.docVIP

三甲益肝颗粒含药血清对HSC-T6细胞形态、增殖和凋亡的影响 杨道坤， 梁海军 ，申保生，孙屹峰 ，乔汉臣 (新乡医学院第一附属医院感染疾病科 ,河南新乡453100) 摘要 目的：观察三甲益肝颗粒含(S-JG)药血清对HSC-T6细胞形态、增殖和凋亡的作用，探讨其抗肝纤维化的细胞作用的机制。方法：不同剂量三甲益肝颗粒灌胃大鼠制备的含药血清，并用不同浓度三甲益肝颗粒含药血清干预HSC-T6细胞株，用倒置相差显微镜观察HSC-T6细胞形态学的变化，MTT法检测S-JG药物血清对HSC-T6细胞增殖的影响，流式细胞仪检测S-JG药物血清诱导HSC-T6细胞早期凋亡。结果：含药血清干预后细胞形态发生明显改变，　MTT实验结果：各组药物血清均可抑制HSC-T6细胞的增殖，S-JG M、H血清组对HSC-T6细胞增殖的抑制均显著高于Sily血清组(P＜0.05)(P＞0.05)effective mechanism of S-JG anti-hepatic fibrosis in cells 【Methods】HSC-T6 cells were exposed in different doses of Sanjiayi gan granula contained serum , The morphous of HSC-T6 line were observed with phase contrast microscope, The proliferation of HSC-T6 were measured by MTT, The apoptosis of HSC-T6 were measured by flow cytometry method.. 【Results】 The morphous changes were observed significantly after treatment with drug sera . The MTT experiment showed that various concentrations of drug sera could inhibit the proliferation of HSC-T6 and the inhibiting level was different. Inhibition ratios of S-JG M drug sera groups and S-JG H drug sera groups were higher than that of Sily drug serum group(P＜0.05). Inhibition ratios of drug sera groups were higher than that of control group(P＜0.05). The floe cytometry method showed that Sanjia yigangranula contained serum can induced the apoptosis of HSC-T6 . And apoptosis ratios was higher withdrug dose’s higher.【Conclusion】Sanjiayi gan granula contained serum can significantly inhibited the proliferation of HSC-T6,induced the apoptosis of HSC-T6. Key words Sanjiayi gan granula HSC-T6 cells Apoptosis Hepatic fibrosis 三甲益肝颗粒是经临床验证治疗肝纤维化有效的中成药复方制剂【1】，我们已经用免疫组化证实三甲益肝颗粒具有通过抑制肝星型细胞活化来抗肝纤维化的作用机制【2】；中药血清药理学研究方法是用含药血清代替煎剂或粗提物进行实验研究的新方法，可避免直接添加药物引起的各种弊端，使中药复方能在细胞分子水平研究其作用机制，其可信性和科学性更强，现将三甲益肝颗粒含药血清抗肝纤维化的实验报道如下。 基金项目：河南省自然科学基金 (No.200733004) 作者简介：杨道坤(1970-) ，男 (汉族) ，主治医师， 硕士. 研究方向：肝硬化的发病机制与治疗. 1材料与方法 1.1 材料 三甲益肝颗粒由本院中药制剂室制成(豫药制，组方：鳖甲、穿山甲、牡蛎、柴胡、大黄、莪术、桃仁、内金、赤芍、丹参、郁金、白术、黄芪、金钱草等，用生理盐水配制成溶液体内给药用。水飞蓟宾胶囊(水林佳)，35mg/粒