【摘要】 采用离体细胞体外孵育法,观察反义c-myb寡脱氧核苷酸（oligodeoxynucletides,ODN）对人绒毛膜促性腺激素（human chori-onic-gonadotropin hormone,hCG）诱导的大鼠间质细胞睾酮分泌的影响,并进一步探讨了外源性二丁酰c-AMP（dbcAMP）、Ca²⁺以及蛋白质抑制剂放线菌酮（cycloheximide,CYX）对间质细胞中c-Myb蛋白表达和睾酮分泌的作用。结果表明,反义c-myb ODN呈剂量依赖性地抑制hCG诱导的离体间质细胞的睾酮分泌,同时使间质细胞中c-Myb蛋白免疫组化染色下降;而无义tat ODN没有相应的作用。100μmol/L的dbcAMP可进一步促使hCG诱导的间质细胞分泌睾酮,并且使间质细胞中c-Myb蛋白免疫组化染色IOD值升高,与hCG组相比,具有统计学意义。钙离子通道阻断剂维拉帕米（10μmol/L）和蛋白质抑制剂放线菌酮（50μg/ml）可使hCG诱导的大鼠间质细胞的睾酮分泌下降,并使间质细胞的C-Myb蛋白兔疫组化染色降低。该结果说明c-myb参与hCG诱导的大鼠间质细胞睾酮分泌作用。更多还原

【Abstract】 The present study was carried out to investigate the effect of antisense c-myb oligodeoxynucleotides （ODN） on hCG-inducedtestosterone secretion in isolated rat Leydig cells. The effects of cAMP, Ca²⁺ and cycloheximide （CYX） on c-Myb protein expression andtestosterone secretion were also observed. The results showed that antisense c-myb ODN inhibited hCG-induced testosterone secretion ofisolated rat Leydig cells in a dose-dependent manner.At the same time, integral optical density immunostaining of Myb in Leydig cells wasalso remarkably reduced. Nonsense tat ODN had no effect on Leydig cells. Further experiments showed that dbcAMP （100μmol/L） obviouslyincreased hCG-induced testosterone secretion and integral optical density （IOD） immunostaining of Myb in Leydig cells. Verapamil （10μmol/L）, a Ca²⁺ channel blocker, and cycloheximide （50μg/ml）, a protein synthesis inhibitor, reduced the immunostaining of c-Myb, and also loweredhCG-induced testosterone secretion in isolated rat Leydig cells. The results indicate that c-myb closely correlates with hCG-induced testoster-one secretion, and that cAMP and Ca²⁺ -dependent pathway participates in the expression of protooncogene.更多还原