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目的观察双氢睾酮对原代培养大鼠Sertoli细胞GDNF及相关细胞信号通路关键蛋白的表达,探讨Sertoli细胞GDNF表达调控的机制。方法原代培养出生后20 d大鼠Sertoli细胞,给予双氢睾酮干预,免疫荧光检测GDNF在细胞中定位和表达强度,Western blot检测GDNF,ERK和CREB蛋白的表达及磷酸化情况,并用ERK和c AMP/PKA信号通路抑制剂干预。结果 GDNF表达在Sertoli细胞质中,双氢睾酮干预可增加GDNF的表达(P<0.05),并使ERK和CREB磷酸化水平增高(P<0.05),使用ERK和c AMP/PKA信号通路抑制剂可减低双氢睾酮诱导的GDNF表达(P<0.05)。结论双氢睾酮通过激活ERK信号通路诱导Sertoli细胞GDNF的表达。
Objective To observe the expression of GDNF and related cell signaling pathways of primary cultured rat Sertoli cells with dihydrotestosterone and to explore the mechanism of GDNF expression in Sertoli cells. Methods Primary sertoli cells were cultured 20 days after birth and treated with dihydrotestosterone. The expression of GDNF in the cells was detected by immunofluorescence. The expressions of GDNF, ERK and CREB were detected by Western blot. The expressions of ERK, c AMP / PKA signaling pathway inhibitor intervention. Results GDNF was expressed in Sertoli cytoplasm. Dihydrotestosterone treatment increased GDNF expression (P <0.05) and increased phosphorylation levels of ERK and CREB (P <0.05). ERK and cAMP / PKA signaling pathway inhibitor Decreased dihydrotestosterone-induced GDNF expression (P <0.05). Conclusion Dihydrotestosterone can induce GDNF expression in Sertoli cells by activating ERK signaling pathway.