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目的探讨促甲状腺激素释放激素受体-1,-2(TRH-R1,TRH-R2)在二甲磺酸乙烷(EDS)处理大鼠睾丸中的表达及其在Leydig细胞中表达的意义。方法构建30只EDS处理大鼠模型,应用蛋白质免疫印迹杂交技术、免疫组织化学ABC法以及免疫荧光双标法,检测TRH-R1和TRH-R2在EDS处理后2d、7d、14d、21d和28d大鼠睾丸中的表达和定位。结果免疫印迹杂交发现,EDS处理后2d至14d,大鼠睾丸组织未见TRH-R1和TRH-R2表达,在EDS处理后21d、28d再次检测到了TRH-R1和TRH-R2的表达;免疫组织化学染色显示,TRH-R1和TRH-R2表达于EDS处理后21d、28d大鼠睾丸生精小管周围的长梭形细胞,免疫荧光双标证实此类细胞为再生的Leydig祖细胞。结论TRH-R1和TRH-R2参与了Leydig细胞的再生过程,可能对成年型Leydig细胞的发育、分化具有调节作用。
Objective To investigate the expression of TRH-R1 and TRH-R2 in the testes of rats treated with ethane dimethylate sulfonate (EDS) and its significance in Leydig cells. Methods Thirty EDS-treated rat models were established. The expressions of TRH-R1 and TRH-R2 were detected by Western blotting, immunohistochemical ABC and immunofluorescence double-labeling method. The expression of TRH-R1 and TRH-R2 were detected at 2d, 7d, 14d, 21d and 28d Expression and localization of rat testes. Results Immunoblotting showed that TRH-R1 and TRH-R2 were not detected in testis of rats from 2 days to 14 days after EDS treatment. The expression of TRH-R1 and TRH-R2 were detected again 21 and 28 days after EDS treatment. Chemical staining showed that TRH-R1 and TRH-R2 were expressed in long spindle cells around the seminiferous tubules of rats on day 21 and 28 after EDS treatment. Immunofluorescence double staining confirmed that these cells were regenerated Leydig progenitor cells. Conclusion TRH-R1 and TRH-R2 are involved in the process of Leydig cell regeneration, which may regulate the development and differentiation of adult Leydig cells.