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Ca^(2+)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Ca^(2+)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Ca^(2+)浓度来完成。
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),50μg/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. Cytosolie 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: Tfi could effectively inhibit the proliferation of LECs in vitro, which may be evoked by changing the concentration of the cytosolic Ca^2+.
出处
《郑州大学学报(医学版)》
CAS
北大核心
2006年第1期130-132,共3页
Journal of Zhengzhou University(Medical Sciences)