CAJ | 학술논문

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.
목적：통과측정DBP대대서고환지지세포내Ca2+、선립체막전위급Caspase활성적영향,탐토DBP시부통과영향선립체막전위이인기지지세포조망.방법건립지지세포원대배양체계,장세포분위0.1% DMSO(대조조)、1 mg/L(저제량조)、10 mg/L (중제량조)、100 mg/L (고제량조),분별처리세포24 h；형광분광광도계측정지지세포내Ca2+적농도,류식세포의검측선립체막전위,매표의측정Caspase-9화Caspase-3적활성.결과여대조조상비,고제량조중Ca2+농도명현승고,차이구유통계학의의(P<0.05),이저、중제량조여대조조비교무통계학의의(P>0.05). DBP도치적선립체막전위승고재중、고제량조중구유통계학의의(P<0.05),수착DBP 농도적증가,Caspase-9여Caspase-3적활성승고,차이균구유통계학의의(P<0.05).결론 DBP능현저강저지지세포선립체막전위,개변선립체막적통투성,진일보승고Caspase-9급Caspase-3적활성,병최종도치고환지지세포조망.
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 <br> 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.