冷应激对人绒毛膜促性腺激素诱导大鼠黄体与颗粒细胞功能的影响

Effect of Cold Stress on Secretory Function of Luteal and Granulose Cells of Rat in vitro

目的为探讨冷应激对大鼠卵巢功能的影响。方法采用体外细胞培养法分离培养大鼠卵巢黄体、颗粒与卵母细胞,设37℃对照组与0、-5、-10、-15、-20、-25℃冷应激组,观察人绒毛膜促性腺激素(hCG)诱导黄体与颗粒细胞分泌孕酮与cAMP的变化,采用放免分析方法测定孕酮与cAMP生成含量,采用四甲基偶氮唑蓝(MTT)法检测细胞存活率。结果-5～-25℃冷应激组黄体细胞和颗粒细胞孕酮含量均低于对照组(均P<0.05),与对照组比较,冷应激组黄体细胞基础cAMP生成量增加,除-20℃组外,差异均有统计学意义(均P<0.05);与对照组比较,除-10℃冷应激组颗粒细胞基础cAMP生成量下降外,其余各组均升高,且差异有统计学意义(P<0.05)。0～-25℃冷应激组细胞存活抑制率呈上升趋势。结论冷应激抑制hCG诱导黄体与颗粒细胞孕酮分泌,协同cAMP生成,使卵母细胞存活率下降,其作用机制有待深入研究。

Objective In order to study the effect of cold stress on the secretory function of rat ovary, the changes of hCG-induced progesterone and cAMP were observed and the livability of the oocytes was determined. Methods The rat luteal and granulose cells and the oocytes were cultured, progesterone and cAMP content was determined by radioimmunoassay kit (RIA kit) under 37 ℃, 0 ℃, -5 ℃, -10 ℃,-15 ℃, -20 ℃ and -25 ℃ respectively. The livability of the oocytes was determined by MTT assay. Results At -5 ℃ to -25 ℃, t...