【作者】 田雄英；

【导师】 林永堃； 翁山耕；

【作者基本信息】 福建医科大学 ， 外科学， 2005， 硕士

【摘要】 目的 研究苦参碱对体外培养的肝星状细胞(HSC)增殖和凋亡的影响,同时观察其对肝细胞有无毒性作用,探讨苦参碱抗肝纤维化的作用机制。 方法 1.体外培养的大鼠肝星状细胞系rHSC-99和肝细胞株HL-7702,以0.125mg/ml、0.25mg/ml、0.5mg/ml的苦参碱处理24小时后,MTT比色法检测细胞增殖。2.体外培养的大鼠肝星状细胞系rHSC-99,以0.25mg/ml的苦参碱处理HSC细胞系rHSC-9924小时后,倒置相差显微镜和透射电镜观察形态改变,TUNEL法原位检测DNA断裂;以0.125mg/ml、0.25mg/ml、0.5mg/ml的苦参碱处理rHSC-9924小时后,Annexin v-FITC/PI双染色流式细胞分析法检测早期细胞凋亡。 结果 1.苦参碱对HSC增殖有明显的抑制作用,且作用呈剂量依赖性(P<0.05);苦参碱对肝细胞增殖有促进作用,但其促增殖作用不随浓度的增加而增强。2.Annexin v-FITC/PI双染色流式细胞分析:对照组、苦参碱0.125mg/ml组、0.25mg/ml组、0.5mg/ml组凋亡率分别是0.99%±0.45%、5.00%±0.98%、13.6%±2.19%、17.2%±2.17%,差异有显著性(F=63.14,P<0.05)。苦参碱处理组细胞TUNEL检测发现DNA断裂,透射电镜观察到苦参碱处理组部分细胞核仁消失,染色质浓缩并凝聚成团块状,沿核膜排列。 结论 苦参碱可抑制HSC增殖、诱导HSC凋亡,可能是苦参碱抗肝纤维化的机制之一。更多还原

【Abstract】 Objective To investigate the effect of matrine on proliferation and apoptosis of Hepatic Stellate Cell in vitro.Methods (1)HSC line,rHSC-99 and hepacyte line,HL-7702 was incubated with matrine of different concertration of 0.125mg/ml , 0.25mg/ml, 0. 5mg/ml for 24h,cell proliferation was assessed by MTT colorimetric assay. (2)rHSC-99 was treated with matrine of different concertration of 0. 125mg/ml, 0.25mg/ml, 0.5mg/ml for 24h, the ealy stage apoptosis was detected with Annexin v-FITC/PI double labbled assay, DNA break was detected by TUNEL in situ assay, morphology of apoptosis was detected by transmission electron microscopy.Results Administration of 0.125-0.5mg/ml matrine could significantly inhibit HSC proliferation in dose-dependent manner (F=14.32, P<0.05) and promot hepacyte proliferation in dose-independent manner(F=86.69 , P<0.05).After treated with matrine at different concertration of 0.125, 0. 25, 0. 5mg/ml matrine for 24h, the apoptosis rate were5. 00%±0. 98% , 13. 6% ± 2.19% , 17. 2% ± 2.17%, respectively, which is significently higher than that of control group(0. 99%±0. 45, F=63.14, P<0.05). DNA break of matrine-treated cells was demonstrated by TUNEL in situ assay, Electron microscopy showed that nucleoli disappeared, chromatin condensted, ranked along inside of nucleonic membrane.Conclusion Matrine can inhibit HSC proliferation and induce apoptosis of HSC, which might be one of the mechanisms of matrine to counteract liver fibrosis.更多还原