双酚A对大鼠睾丸支持细胞糖代谢的影响

发布时间：2018-05-09 09:45

本文选题：双酚A + 支持细胞　；参考：《中华男科学杂志》2015年02期

【摘要】：目的:观察不同浓度双酚A(BPA)对原代培养大鼠睾丸支持细胞(SC)糖代谢及乳酸脱氢酶(LDH)表达的影响,探讨BPA致男性不育的机制。方法:采用两步酶消化法分离雄性Wistar大鼠睾丸SC,建立SC原代培养模型,免疫组化鉴定Fas L。传代后SC随机分为对照组和实验组(100 nmol/L、10μmol/L和1 mmol/L BPA)。培养48 h后,CCK-8法测定细胞增殖活性,核磁共振波谱法测定细胞内代谢物浓度,RT-PCR及Western印迹检测LDH的表达。结果:体外分离培养SC的纯度为(96.05±1.28)%(n=10),CCK-8实验结果显示:与对照组相比,暴露于100 nmol/L BPA组[(98±8)%]、10μmol/L BPA组[(96±3)%]和1 mmol/L BPA组[(95±3)%]的细胞增殖率无明显变化(P0.05);核磁共振波谱显示:与对照组相比,10μmol/L和1 mmol/L BPA作用下SC内葡萄糖及乳酸浓度明显降低(P0.05);RT-PCR及Western印迹结果显示:LDH mRNA的表达随BPA浓度升高呈现降低趋势(100 nmol/L、10μmol/L及1 mmol/L组P均0.05),而LDH蛋白的表达只在1 mmol/L组明显降低(P0.05)。结论:较高浓度BPA降低LDH,影响SC糖代谢过程。推测BPA通过影响SC糖代谢,减少向生殖细胞乳酸的供给,影响精子发生过程。
[Abstract]:Aim: to investigate the effects of different concentrations of bisphenol A (BPA) on glucose metabolism and lactate dehydrogenase (LDH) expression in primary cultured rat testicular Sertoli cells (Sertoli cells) and to explore the mechanism of male infertility induced by BPA. Methods: two step enzyme digestion method was used to isolate male Wistar rat testicular SCS. The primary culture model of SC was established, and Fas L was identified by immunohistochemistry. After passage, SC were randomly divided into two groups: control group and experimental group (10 渭 mol/L and 1 mmol/L BPA). After 48 h culture, the cell proliferation activity was determined by CCK-8 method, the intracellular metabolite concentration was detected by nuclear magnetic resonance spectroscopy (NMR), and the expression of LDH was detected by Western blotting and RT-PCR. Results: the purity of SC isolated and cultured in vitro was 96.05 卤1.28%. The proliferation rate of cells exposed to 10 渭 mol/L BPA group [98 卤8 渭%] 10 渭 mol/L BPA group and 1 mmol/L BPA group [95 卤3 渭%] had no significant change (P 0.05). Nuclear magnetic resonance spectroscopy showed that the concentration of glucose and lactic acid in SC decreased significantly under 10 渭 mol/L and 1 mmol/L BPA compared with the control group. The results showed that the expression of LDH mRNA decreased with the increase of BPA concentration (P < 0.05) in 10 渭 mol/L and 1 mmol/L groups, while the expression of LDH protein in 1 mmol/L group decreased significantly (P 0.05). Conclusion: high concentration of BPA can decrease the level of BPA and affect the process of SC glycometabolism. It is speculated that BPA affects spermatogenesis by affecting the glucose metabolism of SC and reducing the supply of lactic acid to germ cells.
【作者单位】：青岛大学附属医院泌尿外科;济宁医学院附属医院重症医学科;青岛大学附属医院眼科;青岛大学附属医院低温医学科;
【基金】：山东省人口和计划生育委员会科学技术研究项目(201303)~~
【分类号】：R114

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