MEHP抑制小鼠睾丸间质细胞脂噬对睾酮合成的影响  

MEHP Inhibits Lipophagy in Mouse Testicular Interstitial Leydig Cells and Its Effect on Testosterone Synthesis

作　　者：郝羽 李玲[1,2] 闫凤梅 黄静 海云芳 冶月秀 HAO Yu;LI Ling;YAN Fengmei;HUANG Jing;HAI Yunfang;YE Yuexiu(Department of Occupational Hygiene and Environmental Hygiene,School of Public Health,Ningxia Medical University,Yinchuan 750004,China;Ningxia Key Laboratory of Environmental Factors and Chronic Disease Control,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学公共卫生学院职业卫生与环境卫生学系,银川750004 [2]宁夏环境因素与慢性病控制重点实验室,银川750004

出　　处：《宁夏医科大学学报》2023年第12期1194-1200,1221,共8页Journal of Ningxia Medical University

基　　金：宁夏自然科学基金项目(2023AAC03209);宁夏医科大学校级科研基金项目(XM2020008)。

摘　　要：目的探讨邻苯二甲酸单(2-乙基己基)酯[mono(2-ethylhexyl)phthalate,MEHP]对小鼠睾丸间质细胞(TM-3)脂噬水平以及对睾酮合成的影响。方法将对数生长期的TM-3细胞分别暴露于0、200、400、800μmol·L^(-1)的MEHP中,分别为对照组、MEHP低剂量组、MEHP中剂量组、MEHP高剂量组。油红O染色及流式细胞术检测细胞脂滴水平,免疫荧光共定位检测细胞脂噬水平,游离胆固醇检测试剂盒检测细胞中游离胆固醇含量,ELISA检测上清液中的睾酮水平,Western blot检测LC3-Ⅱ/LC3-Ⅰ、P62、SIRT1、FOXO1、Rab7的蛋白表达水平。结果与对照组相比,MEHP低剂量组脂滴含量、睾酮含量、LC3-Ⅱ/LC3-Ⅰ比值及P62蛋白表达水平均无明显变化(P均>0.05);MEHP中、高剂量组脂滴含量、LC3-Ⅱ/LC3-Ⅰ比值及P62均升高(P均<0.05),游离胆固醇含量和睾酮的含量均降低(P均<0.01)。与对照组相比,MEHP低、中、高剂量组TM-3细胞SIRT1、FOXO1、Rab7蛋白表达水平均下降(P均<0.05)。结论MEHP可能通过SIRT1/FOXO1/Rab7信号通路抑制小鼠睾丸间质细胞脂噬,从而减少睾酮含量。Objective To investigate the effect of mono(2-ethylhexyl)phthalate(MEHP)on the level of lipophagy and testosterone synthesis in mouse leydig cells(TM-3)cells.Methods TM-3 cells in the logarithmic growth phase were exposed to 0(control group),200(low dose group),400(medium dose group)and 800(high dose group)μmol·L^(-1) of MEHP.Oil red O staining and flow cytometry were used to detect the level of cell lipid droplets.Immunofluorescence colocalization was used to detect the level of cell lipophagy.Free cholesterol detection kit was applied to detect the free cholesterol content in cells.ELISA was used to detect the level of testosterone in the supernatant.Western blot was used to detect the protein expression levels of LC3-Ⅱ/LC3-Ⅰ,P62,SIRT1,FOXO1 and Rab7.Results Compared with the control group,there were no significant changes in lipid drop content,testosterone content,LC3-Ⅱ/LC3-Ⅰand P62 protein expression levels in the MEHP low dose group(P all>0.05),lipid droplet content,LC3-Ⅱ/LC3-Ⅰand P62 increased(P all<0.05)in the MEHP medium and high dose groups,and free cholesterol content and testosterone content decreased(P all<0.01).Compared with the control group,the expression levels of SIRT1,FOXO1 and Rab7 proteins in TM-3 cells in the low,medium and high dose groups were decreased(P all<0.05).Conclusion MEHP may inhibit lipophagy in mouse Leydig cells through SIRT1/FOXO1/Rab7 signaling pathway and reduce testosterone content.

关 键 词：脂噬 邻苯二甲酸单(2-乙基己基)酯 小鼠睾丸间质细胞 睾酮 

分 类 号：R114[医药卫生—卫生毒理学]