大鼠卵巢蒂扭转后颗粒细胞凋亡及相关蛋白表达

Granulosa Cell Apoptosis and Related Proteins Expression in Experimental Torsed Ovaries of the Adult Female Rats

目的探讨大鼠卵巢蒂扭转后不同时间颗粒细胞的凋亡及与Bax和Caspase-3蛋白表达的关系,用实验数据验证卵巢蒂扭转后保守性手术治疗的最佳时间.方法将20只Sprague-Dawley性成熟雌性大鼠按照随机设计原则分成4组,每组均为5只.此4组分为正常组,卵巢蒂扭转后24h,48h,72h组.取卵巢组织常规石蜡包埋,切片,HE染色,采用免疫组化(SP)法检测Bax和Caspase-3及(TUNEL)测定颗粒细胞凋亡.SPSS进行统计分析,P<0.05为差异有统计学意义.结果SD性成熟雌性大鼠在卵巢扭转后,TUNEL检测细胞凋亡在第48h达高峰,与对照组比较,在第24h、48h、72h时均有统计学意义(P<0.05).实验组Bax的表达随扭转时间的增加而表达增加,扭转后第72h达高峰.与对照组比较,实验组Bax的表达在卵巢扭转后第24h、48h、72h均有统计学意义(P<0.05).Caspase-3蛋白在对照组几乎无表达,实验组Caspase-3的表达随扭转天数的增加而增加,扭转后第72小时达高峰.与对照组比较,实验组Caspase-3的表达在卵巢扭转后第24h、48h、72h均有统计学意义(P<0.05).Bax和Caspase-3蛋白在卵巢扭转后细胞凋亡的表达中呈正相关.结论卵巢扭转后可诱导颗粒细胞发生凋亡,其机制与Bax和caspase-3的激活有关.同时根据TUNEL检测结果提示,卵巢扭转后,复位卵巢并保留卵巢的保守性手术治疗的最佳时机是扭转48h内,而且时间越早越好.

Objective To explore granulosa cells apoptosis and its relationship with Bax nd Caspase-3 proteint expression at different times in experimental torsed ovaries of the adult female rats, and to validate the best time of conservative surgical treatment of ovarian torsion. Methods Twenty adult female Spragne-Dawley rats were divided randomly into four groups （n = 5 in each group） ： instantaneous control, following an ovarian torsion period of 24 h, 48 h, 72 h. All removed ovarian tissues were embedded in paraffin, cut into sections, and stained with Hematoxylin-eosin. Immunohistochemic technical was applied to investigate the changes of expression of Bax and Caspase-3 in ovarian tissues. The evidence of granulosa cell apoptosis was detected in these tissues by TUNEL. SPSS software was used to analyze the data, and P 〈 0.05 was considered the level of significance. Results The apoptotic cell count of TUNEL in torsion group was peaked at 48 h after fight ovarian torsion in adult female rats, demonstrating significant differences at 24 h, 48 h, 72 h compared with that observed in the control group （P 〈 0.05）. With the time of ovarian torsion, Bax protein expression in the experimental group was up-regulated and peaked at 72 h, demonstrating significant differences at 24 h, 48 h and 72 h compared with the control group （P 〈 0.05 ）. Caspase-3 protein was hardly found in the ovarian tissue of control group. With the time of ovarian torsion, Caspase-3 protein expression in the torsion group was up-regulated and peaked at 72 h, demonstrating significant differences at 24 h, 48 h and 72h compared with the control group （ P 〈 0.05 ）. The expression of Bax and Caspase-3 protein had a positive correlation with cell apoptosis after ovarian torsion. Conclusions The mechanisms of granulosa cell apoptosis is highly associated with the activation of Bax and Caspase-3 in experimental ovarian torsion. According to the results of TUNEL, the best time is within 24 hours to treat adnexal torsion conservatively, and the sooner the better.