激光共聚焦检测双酚A致精母细胞损伤后线粒体膜电位的变化

发布时间：2018-02-14 10:52

本文关键词： 激光扫描共聚焦显微镜双酚A 精母细胞线粒体膜电位荧光成像荧光探针JC-　出处：《中国医学装备》2016年02期 　论文类型：期刊论文

【摘要】：目的:运用激光扫描共聚焦显微镜(LSCM)技术检测双酚A(BPA)致精母细胞损伤后线粒体膜电位的变化,探讨其作用机制。方法:将培养的GC-2小鼠精母细胞分为对照组、低剂量组和高剂量组,分别加入0、10μmol/L、100μmol/L的BPA,一同培养3 h后使用荧光探针JC-1对3个组样本分别进行标记,采用LSCM检测胞内线粒体内JC-1荧光强度的变化。利用LSCM专用软件分析荧光强度,通过荧光颜色的转变检测线粒体膜电位的变化,用红绿荧光的相对比例衡量线粒体去极化的比例。结果:对照组、低剂量组和高剂量组之间红绿荧光的比值有着显著差异。BPA低剂量组和高剂量组的胞内线粒体膜电位明显低于对照组,而高剂量组的胞内线粒体膜电位较低剂量组低。结论:BPA对精母细胞有损伤,且随着剂量加大,损伤越严重。LSCM技术方法灵敏度高,能够实时监测细胞内线粒体膜电位的变化。
[Abstract]:Objective: to investigate the changes of mitochondrial membrane potential (MMP) after spermatocyte injury induced by bisphenol A (BPA) by laser scanning confocal microscopy (LSCM). Methods: the cultured spermatocytes of GC-2 mice were divided into control group. In the low dose group and the high dose group, 10 渭 mol 路L ~ (-1) L ~ (-1) of 0 渭 mol / L ~ (100 渭 mol/L) was added respectively. The three groups were labeled with fluorescent probe JC-1 for 3 h. The changes of JC-1 fluorescence intensity in mitochondria were detected by LSCM, the fluorescence intensity was analyzed by LSCM software, and the change of mitochondrial membrane potential was detected by fluorescence color transformation. The ratio of mitochondrial depolarization was measured by the relative ratio of red and green fluorescence. Results: control group, There was significant difference in the ratio of red and green fluorescence between the low dose group and the high dose group. The intracellular mitochondrial membrane potential of BPA low dose group and high dose group was significantly lower than that of the control group. Conclusion: BPA has damage to spermatocytes, and with the increase of dose, the damage is more serious. LSCM technique has high sensitivity and can monitor the changes of mitochondrial membrane potential in the cells in real time. [WT5 "HZ] [WT5" BZ] [WT5BZ] [WT5 "BZ] [WT5" BZ] [WT5 "BZ]
【作者单位】：第三军医大学生物医学分析测试中心;
【分类号】：R114

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