DBP对大鼠睾丸支持细胞内Ca^(2+)、线粒体膜电位及Caspase活性的影响

Effects of DBP on Intracellular Ca^(2+),Mitochondrial Membrane Potential and Caspase Activity in Sertoli Cells in Rats

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(P<0.05),而低、中剂量组与对照组比较无统计学意义(P>0.05).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(P<0.05),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(P<0.05).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.

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.