标题：非酶糖基化终末产物介导的ERK1/2信号转导通路在大鼠睾丸间质细胞睾酮分泌中的作用

      作者：戚亚伟 1，胡传银 2，陈绍红 3，赵云涛 3，刘铀 3    (1.广东医科大学附属医院整形外科研究所，广东 湛江 524001；2.广东医科大学细胞生物学教研室，广东 湛江 524023；3.广东海洋大学现代生化中心，广东 湛江 524088)?

      卷次： 2017年28卷1期

      【摘要】 目的 探讨非酶糖基化终末产物(AGEs)对大鼠睾丸间质细胞睾酮分泌的影响及其潜在机制。方法 首先，原代培养大鼠睾丸间质细胞，采用3β-HSD免疫细胞化学方法对间质细胞进行鉴定；其次，MTT法测定不同浓度的AGEs (25 μg/mL、50 μg/mL、100 μg/mL、200 μg/mL)及200 μg/mL BSA作用Leydig细胞后的细胞活力；最后，用不同浓度的AGEs及200 μg/mL BSA预处理Leydig细胞12 h，之后更换含相应浓度AGEs或BSA的培养基并加入终浓度为4 U/mL的hCG，其中对照组不含AGEs和BSA，ELISA法和Western blot分别检测睾酮分泌量和p-ERK1/2的表达量。结果 MTT法表明AGEs (≤200 μg/mL)对Leydig细胞的活力在本实验所研究的时间内没有影响；ELISA法和Western blot结果显示，不同浓度AGEs处理后，hCG诱导的 Leydig细胞睾酮合成量和ERK1/2蛋白的磷酸化都呈现浓度依赖性下降，与对照组相比，差异有统计学意义(P<0.01)。结论 AGEs通过抑制ERK1/2的磷酸化降低大鼠Leydig cells睾酮的分泌。
      【关键词】 非酶糖基化终产物；睾丸间质细胞细胞；细胞外调节蛋白激酶；睾酮
      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2017）01—0001—05

Role of ERK1/2 signaling pathway mediated by advanced glycation end products in testosterone secretion of ratLeydig cells.
QI Ya-wei 1, HU Chuan-yin 2, CHEN Shao-hong 3, ZHAO Yun-tao 3, LIU You 3. 1. Institute of PlasticSurgery, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, Guangdong, CHINA; 2. Department ofCell Biology, Guangdong Medical University, Zhanjiang 524023, Guangdong, CHINA; 3. Modern Biochemistry Center,Guangdong Ocean University, Zhanjiang 524088, Guangdong, CHINA
【Abstract】 Objective To investigate the effect of advanced glycation end products (AGEs) on testosterone pro-duction of rat Leydig cells and its underlying mechanism. Methods First, the primary Leydig cells were isolated, purifiedand cultured in vitro. The stromal cells were identified by 3β-HSD immunocytochemistry. Second, the effects of variousconcentrations of AGEs (25 μg/mL, 50 μg/mL, 100 μg/mL and 200 μg/mL) and BSA (200 μg/mL) on Leydig cell viabilitywere evaluated by MTT. Finally, Leydig cells were pre-incubated with various concentrations of AGEs and 200 μg/mLBSA for 12 h. Then, the culture medium was replaced with fresh medium containing human chorionic gonadotropin(hCG, 4 U/mL) with or without the same concentrations AGEs and BSA. The control group did not contain AGEs andBSA. The secretion of testosterone induced by human chorionic gonadotropin (hCG) was determined by ELISA. The pro-tein expression levels of ERK1/2 phosphorylationwere were measured by western blot. Results MTT data indicated thatthe viability of the Leydig cells treated with AGEs (concentrations≤200 μg/mL) was not significantly altered. ELISA andwestern blot showed that AGEs could remarkably suppress testosterone production and decrease the phosphorylation levelsof p-ERK1/2 induced by hCG in a concentration-dependent manner in rat Leydig cells compared with the control group(P<0.01). Conclusion AGEs decrease testosterone secretion in rat Leydig cells by inhibiting ERK1/2 phosphorylation.
      【Key words】 Advanced glycation end products (AGEs); Leydig cell; Extracellular signal-regulated kinase 1/2(ERK1/2); Testosterone·论 著·