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目的研究高血糖对排卵前小鼠卵母细胞发育及颗粒细胞凋亡的影响。方法出生后20d ICR雌鼠,建立急性高血糖模型;取高血糖模型雌鼠和正常雌鼠超排卵,分别于hCG注射后的0、2、6和10h取卵巢组织,石蜡切片,TUNEL检测凋亡,免疫组织化学ABC法检测连接蛋白connexin-43的表达,相同时间点取生发泡期(GV期)卵母细胞统计生发泡破裂(GVBD)和第一极体(FB1)的发生率。结果1.高血糖组卵巢湿重与小鼠体重的比值与排卵数目均低于正常组(P<0.01);2.免疫组织化学显示,hCG注射后6h、10h颗粒细胞表达connexin-43在高血糖组明显低于正常组(P<0.05);3.高血糖组卵泡颗粒细胞的凋亡率明显高于正常组,hCG注射后6h、10h差异最显著(P<0.05);4.高血糖组卵母细胞GVBD发生率低于正常组,于注射后2h差异最显著(P<0.01)。结论高血糖环境下卵母细胞发育延迟,颗粒细胞的凋亡率增高。这种发育延迟与颗粒细胞间connexin-43低表达呈正相关。
Objective To study the effects of hyperglycemia on the development of oocytes and the apoptosis of granulosa cells in mice before ovulation. Methods ICR female rats were sacrificed 20 days after birth to establish an acute hyperglycemic model. Ovarian tissues were obtained at 0, 2, 6 and 10 h after hCG injection in ovariectomized female rats and normal female rats. Paraffin sections and TUNEL assay The immunohistochemical ABC method was used to detect the expression of connexin-43 and the incidence of GVBD and the first polar body (FB1) in the oocytes at the same time point in the germinal foaming stage (GV stage). Results 1. The ratio of ovarian wet weight to body weight and the number of ovulation in hyperglycemia group were lower than those in normal group (P <0.01) .2. Immunohistochemistry showed that the expression of connexin-43 at 6h and 10h after hCG injection was significantly higher (P <0.05). 3. The apoptosis rate of granulosa cells in hyperglycemia group was significantly higher than that in normal group (P <0.05) at 6h and 10h after injection of hCG (P <0.05) The incidence of GVBD in group oocytes was lower than that in normal group at 2h after injection (P <0.01). Conclusion Oocyte development is delayed and the rate of apoptosis of granulosa cells is increased in hyperglycemic environment. This developmental delay was positively correlated with the low expression of connexin-43 in granulosa cells.