反义c-fos寡脱氧核苷酸对hCG诱导大鼠黄体细胞孕酮和雌二醇生成的影响

EFFECT OF C-FOS ANTISENSE OLIGODEOXYNUCLEOTIDE ON THE HCG-INDUCED PROGESTERONE AND ESTRADIOL PRODUCTION IN RAT LUTEAL CELLS

采用离体孵育大鼠黄体细胞的方法，观察了反义c-fos寡脱氧核苷酸（反义c－fosODN）对hCG诱导的黄体细胞孕酮（P）和雌二醇（E2）产生的影响，同时观察了外源性cAMP和钙离子通道阻断剂维拉帕米（Verapamil）对黄体细胞中c－fos蛋白的影响。结果发现，反义c－fosODN能呈剂量相关方式抑制hCG诱导的黄体细胞P和E2的产生，同时使c－fos蛋白染色阳性的黄体细胞百分数下降；而无义tatODN没有相应的作用。10-4mol／L二丁酰cAMP能明显增强hCG对P、E2产生的刺激作用，并使c－fos蛋白染色阳性的黄体细胞百分率显著升高；当与反义c－fosODN共同作用时能明显逆转反义c-fosODN对P、E2产生和c－fos表达的抑制作用。10～6mol／L维拉帕米（Verapamil）显著抑制hCG诱导的P、E2产生，同时使c－fos蛋白染色阳性的黄体细胞百分率下降；当与反义c－fosODN共同作用时，对反义c-fosODN对P、E2产生和c－fos表达的抑制作用具有协同效应。表明，c－fos与hCG诱导的黄体细胞孕酮和雌二醇的产生密切相关；cAMP和Ca2+能调节c－fos在黄体细胞中的表达。

The effects of c-fos antisense oligodeoxynucleotides (antisense c-fos ODN) on the hCG-induced progesterone and estradiol production in isolated rat luteal cells and the effect of cAMP and Ca2+on c-fos oncoprotein in luteal cells were investigated. Antisense cfos ODN inhibited the hCG-induced progesterone and estradiol production in a dose-dependent manner, and the percentages of luteal cells stained for c-fos protein were also decreased significantly. Nonsense tat ODN had no similar effects on luteal cells. Further experiments showed that 10-4mol/L dbcAMP increased the percentage of luteal cells stained for c-fos protein,and increased the hCG-induced P and E, production. When combined with antisense c-fos ODN, dbcAMP reversed the inhibition of hCG-induced P and E, production, and increased the percentage of luteal cells stained for c-fos protein. However verapamil (10-6mol/L) had opposite effects on c-fos expression and hCG-induced P and E, production as compared with dbcAMP.