氯丙嗪对大鼠卵巢颗粒细胞损伤及其作用机制

The damage of ovarian granulosa cells in rats in vitro caused by chlorpromazine and its mechanism

目的通过氯丙嗪染毒体外培养的大鼠卵巢颗粒细胞,研究其对卵巢颗粒细胞的毒性作用,并初步探讨可能的作用机制。方法对未成熟的Wistar大鼠卵巢颗粒细胞进行原代培养,用不同浓度的氯丙嗪(0、0.1、1和10μmol/L)染毒24 h。MTT法检测细胞相对活力,ELISA法检测培养液中孕酮(P)和雌二醇(E2)的含量,Hoechst 33258检测颗粒细胞的凋亡变化。结果在本次实验所设置的剂量范围内,与对照组比较,氯丙嗪能明显地抑制大鼠卵巢颗粒细胞增殖和E2的分泌(P<0.05),促进颗粒细胞凋亡(P<0.01),呈浓度依赖关系;而低剂量的氯丙嗪(0.1μmol/L)却能够促进P的分泌(P<0.05),但是随着剂量的升高,P的分泌量逐渐下降。结论氯丙嗪可通过抑制卵巢颗粒细胞增值、诱导颗粒细胞凋亡并影响细胞甾体激素的分泌,而引起卵巢功能障碍。

Objective To study the toxic effects and explore the possible mechanism of Chlorpromazine exposure to rat ovarian granulosa cells in vitro.Methods Immature primary cultured ovarian granulosa cells of wister rat were treated with various concentrations of chlorpromazine（0,0.1,1,10 μmol/L） for 24 h.After 24-h exposure to chlorpromazine,cell viability was detected by MTT assay,content of progesterone（P） and estradiol（E2） collected from medium were detected by ELISA assay.And ovarian granulosa cells stained with Hochst dye 33258 were checked for apoptotis.Results Compared to the control group,chlorpromazine could inhibit the proliferation of ovarian granulosa cells and the secretion of E2（P 0.05） and promote the apoptosis of granulosa cells（P 0.01） in a concentration dependent manner.But progesterone secretion increased at low-dose chlorpromazine（P 0.05）.And the progesterone secretion decreased following the increased concentrations of chlorpromazine.Conclusion Chlorpromazine could significantly decreased the cell viability of rat ovarian granulosa cells,induced apoptosis and affected secretion of E2 and P,which resulted in ovarian dysfunction.