【摘要】 目的:探讨雷公藤内酯醇（Tri）对牛晶状体上皮细胞（lens epithelial cell,LECs）增殖的影响及可能的机制。方法:取体外培养的第4代牛LECs,分5组培养。空白对照组:以DMEM培养;增殖对照组:以DMEM+50μg/L rhEGF培养;Tri高、中、低剂量组分别以含50μg/L rhEGF和40μg/L、20μg/L、10μg/L Tri的DMEM培养。分组培养6h、12h、24h、48h、72h后用MTT比色法及流式细胞仪检测细胞增殖抑制率和增殖细胞核抗原（PCNA）阳性表达率;用荧光指示剂Fura-2/AM测定空白对照组、增殖对照组、Tri中剂量组分组培养48h后胞内游离Ca²⁺浓度。结果:随着作用时间的延长和浓度的增加,Tri对rhEGF诱导的LECs增殖的抑制率逐渐增大;Tri各剂量组PCNA阳性表达率低于增殖对照组;Tri中剂量组胞内Ca²⁺浓度高于增殖对照组和空白对照组。结论:Tri对LECs的增殖有明显的抑制作用,该作用可能通过影响胞内Ca²⁺浓度来完成。更多还原

【Abstract】 Aim: To explore the effects and the mechanism of triptolide (Tri) on the proliferation of cultured bovine lens epithelial cells (LECs) in vitro. Methods: The fourth generation of bovine LECs were allocated into 5 groups and cultured with DMEM (normal control group), DMEM +50 μg/L rhEGF (proliferation group),50 μg/L rhEGF+40 μg/L Tri +DMEM (high-dose Tri group),50 μg/L rhEGF+20 μg/L Tri +DMEM (medium-dose Tri group),50 μg/L rhEGF+ 10 μg/L Tri+DMEM (high-dose Tri group).The inhibition rates of LECs and the expression rates of proliferating cell nuclear antigen (PCNA) were measured by MTT (colorimetric assay) and FCM(flow cytometry)at 6 h,12 h,24 h,48 h,and 72 h after culture,respectively. Cytosolic Ca~ 2+ was determined by the fluorescence determination with Fura-2 at 48 h after culture. Results: The inhibition rates of LECs changed in the time-dose-dependent way.In low-dose, medium-dose and high dose Tri groups, the positive rate of PCNA was lower than that of proliferation group. The intracellular Ca~ 2+ in medium-dose Tri group was higher than those in proliferation group and normal control group.Conclusion: Tri could effectively inhibit the proliferation of LECs in vitro, which may be evoked by changing the concentration of the cytosolic Ca~ 2+ .更多还原