摘要
目的观察蛋白酶体抑制剂MG132对诱导体外培养激活的肝星状细胞(HSC)凋亡的影响。方法大鼠肝星状细胞分离采用胶原酶原位灌注法,用流式细胞仪和琼脂糖凝胶电泳法检测MG132对诱导激活的HSC凋亡的影响。结果1、2、3μmol/LMG132培养HSC24h后,细胞周期分析发现S期细胞减少,G2/M期细胞显著增加(P<0.01),呈现一个剂量依赖性的关系;流式细胞术检测到明显的亚G1峰,各组的凋亡指数(%)分别是12.70±1.7、17.52±2.3、22.60±3.4,与对照组(1.9±0.6)相比,差异有显著性(P<0.01);3μmol/LMG132作用12、24、36、48h,凋亡指数(%)分别是16.43±2.2、22.60±2.7、29.80±1.7和36.30±1.4,与对照组相比,差异有显著性(P<0.01),呈现一个时间依赖性的关系;琼脂糖凝胶电泳可以看到明显的DNA梯带的形成。结论MG132能够诱导激活的HSC发生凋亡,且在发生凋亡之前有一个明显的G2/M期的阻滞。
Objective To investigate the effect of MG 132 on apoptosis of the activated rat hepatic stellate cells（HSC）. Methods HSC were isolated from rats with collagenase in situ liver recirculating perfusion. Apoptosis was analyzed by flow cytometry（ FCM ） and agarose get electroporesis. Results Admilistrition of 1 -3 μmol/L MG 132 could significantly inhibit HSC proliferation in dose dependent manner（ P 〈0. 01 ）. After treated with MG 132 in all different concentration of 1 - 3 μmoL/l, for 24 h,the number of in G2/M phase increased and in S phase decreased. There was significant sub G1 peak using FCM. Apoptosis indexes（ % ） were 12.70 ± 1.7,17. 52 ±2.3,22.60 ± 3.4 respectively, which were significantly higher than thai of control group （ 1.9 ±0. 6 ,P 〈0.01 ）. Ineubated with MG 132 in for 12,24,36 and 48 h at the same concertration of 3 μmol/L,apoptosis indexes（ % ） were 16.43 ±2.2,22. 60 ±2.7,29.80 ± 1.7 and 36.30 ±1.4 respcetively, which were significantly higher than that of control group,and it shows relationship of time dependence. An oligonucleosomal DNA ladder of MG 132 - treated cells was demonstrated, Conclusion Proteasome inhibitor MG 132 can induce apoptosis of HSC and can induce G2/M arrest before the apoptosis appeared in HSC cell.
出处
《临床消化病杂志》
2005年第6期267-269,共3页
Chinese Journal of Clinical Gastroenterology
