DBP对大鼠睾丸支持细胞内Ca2+、线粒体膜电位及Caspase活性的影响

目的通过测定DBP对大鼠睾丸支持细胞内Ca2+、线粒体膜电位及Caspase活性的影响,探讨DBP是否通过影响线粒体膜电位而引起支持细胞凋亡.方法建立支持细胞原代培养体系,将细胞分为0.1%DMSO(对照组)、1 mg/L(低剂量组)、10 mg/L(中剂量组)、100 mg/L(高剂量组),分别处理细胞24 h;荧光分光光度计测定支持细胞内Ca2+的浓度,流式细胞仪检测线粒体膜电位,酶标仪测定Caspase-9和Caspase-3的活性.结果与对照组相比,高剂量组中Ca2+浓度明显升高,差异具有统计学意义(P0.05),而低、中剂量组与对照组比较无统计学意义(P0.05).DBP导致的线粒体膜电位升高在中、高剂量组中具有统计学意义(P0.05),随着DBP浓度的增加,Caspase-9与Caspase-3的活性升高,差异均具有统计学意义(P0.05).结论 DBP能显著降低支持细胞线粒体膜电位,改变线粒体膜的通透性,进一步升高Caspase-9及Caspase-3的活性,并最终导致睾丸支持细胞凋亡.

Effects of DBP on Intracellular Ca2+,Mitochondrial Membrane Potential and Caspase Activity in Sertoli Cells in Rats

Objective To explore the mechanism of DBP induced sertoli cell apoptosis by determining the effects of DBP on the concentration of Ca2+,the mitochondrial membrane potential and the activities of Capase in sertoli cells in rats. Method The primary cultivation system was established, and the sertoli cells were divided into 0. 1% DMSO (negative control),1 mg/L DBP (low dose),10 mg/L DBP (medium dose) and 100 mg/L DBP ( high dose ) . After culturing for 24 h, the concentrations of Ca2+ in the sertoli cells were detected by fluorospectrophotometer,a flow cytometer was used to detect the changes of mitochondrial membrane potential, and microplate reader was applied to measure the activities of Caspase-9 and Caspase-3. Results Compared with the control group,there was significant increase of Ca2+ in the high dose group(P<0. 05),while the changes in the low and the medium dose groups were unremarkable ( P>0 . 05 ) . The impairment caused by DBP had statistical significance in the medium and the high dose groups ( P<0 . 05 ) . With the increase of DBP , the
activities of Caspase-9 and Caspase-3 were all improved significantly ( P<0 . 05 ) . Conclusion DBP could decrease the mitochondrial membrane potential,change the permeability of mitochondrial membrane,and further increase the activities of Caspase-9 and Caspase-3,at last lead to sertoli cells apoptosis.