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目的通过对原代培养的大鼠睾丸支持细胞进行邻苯二甲酸二乙基己酯(di-2-ethylhexyl phthalate,DEHP)染毒,探讨DEHP体外对大鼠睾丸支持细胞间隙连接蛋白43(connexin 43,Cx43)表达及间隙连接通讯(gap junction intercellularcommunication,GJIC)功能的影响。方法体外分离和原代培养大鼠睾丸支持细胞,应用不同浓度的DEHP(10、50、100nmol/L)作用于支持细胞24 h,应用荧光定量PCR法检测Cx43基因mRNA表达,用Weastern blot方法检测Cx43蛋白表达,划痕标记荧光染料示踪方法检测GJIC功能。结果与对照组相比,DEHP各组支持细胞Cx43基因mRNA和蛋白表达均下降(P<0.05),呈浓度依赖关系,同时支持细胞GJIC功能降低。结论 DEHP在体外可通过抑制大鼠睾丸支持细胞Cx43基因表达而抑制GJIC功能,进而影响支持细胞功能。
OBJECTIVE: To investigate the effects of DEHP on the connexin 43 (connexin 43) in rat primary cultured rat sertoli cells by di-2-ethylhexyl phthalate (DEHP) 43, Cx43) expression and gap junction intercellularcommunication (GJIC) function. Methods Sertoli cells were isolated and cultured in vitro. The cells were treated with various concentrations of DEHP (10, 50 and 100 nmol / L) for 24 h. The mRNA expression of Cx43 was detected by fluorescence quantitative PCR and detected by Weastern blot Cx43 protein expression, scratch mark fluorescent dye tracer method to detect GJIC function. Results Compared with the control group, the mRNA and protein expressions of Cx43 in all the DEHP groups were decreased (P <0.05) in a concentration - dependent manner, and the function of GJIC was also decreased. Conclusion DEHP can inhibit GJIC function in vitro by inhibiting the expression of Cx43 gene in rat testicular support cells, thereby affecting the function of supporting cells.