雷公藤内酯醇抑制牛晶状体上皮细胞增殖

目的探讨雷公藤内酯醇(Triptolide,Tri)对体外培养的牛晶状体上皮细胞(bovine Lens epithelial cell,BLECs)增殖的抑制作用。方法取第4代对数生长期的牛LECs,培养24 h后,加入重组人表皮生长因子(recombi-nant human epithelial growth factor,rhEGF)(终浓度50μg/L)和不同浓度的Tri(40、20、10μg/L),再分别作用6、12、24、48、72 h后,采用MTT法及流式细胞仪检测增殖细胞核抗原(PCNA)在LECs的阳性表达率和增殖抑制率,观察Tri对LECs增殖的抑制作用。结果不同浓度Tri均可抑制处于增殖状态的LECs,并呈剂量和时间依赖性;72 h增殖抑制率分别达到80.09%、61.29%及39.93%。PCNA表达率在6-72 h内随浓度增高和作用时间延长有明显的下调,呈明显的时间-效应关系和剂量-效应关系,且各浓度Tri组与同一时点增殖对照组相比均有非常显著性差异(P<0.01)。结论Tri对牛晶状体上皮细胞增殖有明显的抑制作用。

Experimental studies of triptolide inhibition on the proliferation of lens epithelial cells

Objective To determine the effects and mechanisms of the triptolide inhibition on the proliferation of bovine lens epithelial cells(BLECs) in vitro.Methods The fourth passage of BLECs incubated for 24 hours were cultured with the recombinant human epithelial growth factor(rhEGF) of 50μg/L and triptolide in different concentrations(40,20,10μg/L) for 6-72 hours(6h,12h,24h,48h and 72h).The inhibition rates of BLECs and the expression rates of proliferating cell nuclear antigen(PCNA) were measured by MTT(colorimetric assay) and FCM(flow cytometry),in order to observe and determine the inhibition effects of triptolide on the proliferation of BLECs.Results With the different concentrations of triptolide(40, 20 and (10μg/L)),the inhibition rates of proliferation of BLECs in vitro changed in a time-dose-dependent way.The inhibition rates for 72h were 80.09%,61.29% and 39.93%,respectively.The expressions of PCNA(6-72h) changed with the higher concentrations and the longer time length in a down-regulating way,and compared with other control groups of the same time length,there were significant differences(P<0.01).The time-dose-dependent relations were evident.Conclusion Triptolide effectively inhibits the proliferation of BLECs in vitro.