育肾助孕方对卵巢储备功能降低大鼠卵巢Bcl-2、Bax、Caspase-3表达的影响

Effects of Yushen Zhuyun decoction on expression of Bcl-2, Bax and Caspase-3 in ovaries of rats with diminished ovarian reserve

目的观察育肾助孕方对卵巢储备功能降低模型大鼠卵巢Bcl-2、Bax、Caspase-3 mRNA及蛋白表达的影响。方法48只雌性SD大鼠随机分为对照组、模型组、补佳乐组及育肾助孕方低、中、高剂量组,每组8只。除对照组外,其余各组均采用雷公藤多苷药液灌胃建立卵巢储备功能降低大鼠模型。各组予相应药物干预2周。采用免疫组化及Western blot检测卵巢组织细胞中Bcl-2、Bax、Caspase-3蛋白表达情况,采用Real-time PCR检测卵巢细胞中Bcl-2、Bax、Caspase-3 mRNA表达情况。结果与对照组比较,模型组Bcl-2 mRNA及蛋白表达显著降低,Bax、Caspase-3 mRNA及蛋白表达显著升高(P<0.05)。与模型组比较,补佳乐组和育肾助孕方各剂量组Bcl-2蛋白表达显著升高,Bax、Caspase-3 mRNA及蛋白表达显著降低(P<0.05);补佳乐组和育肾助孕方高剂量组Bcl-2 mRNA表达显著高于模型组(P<0.05)。与育肾助孕方低剂量组比较,育肾助孕方高剂量组Bcl-2 mRNA表达显著升高,育肾助孕方高中剂量组Bax蛋白表达明显降低(P<0.05)。结论育肾助孕方可通过上调卵巢组织细胞中Bcl-2 mRNA及蛋白表达,下调Bax、Caspase-3 mRNA及蛋白表达,促进卵巢颗粒细胞增殖,抑制其过度凋亡,从而改善卵巢储备功能。

Objective To investigate the effects of Yushen Zhuyun decoction on the protein and mRNA expression of Bcl-2,Bax and Caspase-3 in ovaries of rats with diminished ovarian reserve. Methods Forty-eight female SD rats were randomly divided into control group,model group,Prognova group,and low-,medium-and high-dose Yushen Zhuyun decoction groups,8 rats in each group. Except for the control group,the other groups were treated with tripterygium glycosides to establish rat models of diminished ovarian reserve. All of the animals were treated for two weeks. The protein expression levels of Bcl-2,Bax and Caspase-3 in ovarian tissues and cells were detected by immunohistochemistry and Western blot. The mRNA expression levels of Bcl-2,Bax and Caspase-3 in ovarian cells were detected by Real-time PCR. Results Compared with the control group,the protein and mRNA expression levels of Bcl-2 decreased significantly,and the protein and mRNA levels of Bax、Caspase-3 increased siginifantly in the model group( P<0.05). Compared with the model group,the protein expression level of Bcl-2 increased significantly,and the protein and mRNA expression levels of Bax and Caspase-3 decreased significantly in the Prognova group and the Yushen Zhuyun decoction groups( P<0.05). The mRNA expression levels of Bcl-2 in the Prognova group and the high-dose Yushen Zhuyun decoction group was significantly higher than that in the model group( P<0.05). Compared with the low-dose Yushen Zhuyun decoction group,the mRNA expression level of Bcl-2 increased significantly in the nigh-dose Yushen Zhuyun decoction group,and the protein expression level of Bax decreased obviously in the medium-and high-dose Yushen Zhuyun decoction group( P<0.05). Conclusion Yushen Zhuyun decoction can up-regulate the protein and mRNA expression of Bcl-2,down-regulate the protein and mRNA expression of Bax and Caspase-3 in ovarian tissues and cells to promote proliferation and inhibit excessive apoptosis of ovarian granulosa cells,thus improving the function of ovarian reserve.