补肾养精颗粒调节早发性卵巢功能不全模型大鼠卵泡颗粒细胞凋亡机制研究

Mechanism of Bushen Yangjing Granules in Regulating Follicular Granulosa Cell Apoptosis of Model Rats with Premature Ovarian Insufficiency

目的:探讨补肾养精颗粒调节早发性卵巢功能不全(POI)模型大鼠卵泡颗粒细胞凋亡的机制。方法:60只通过雷公藤多苷片造模POI大鼠,随机分为补肾养精颗粒高、中、低剂量组,空白组及非模型组。干预组给予不同剂量补肾养精颗粒连续灌胃15 d,空白组及非模型组给予生理盐水灌胃。连续观测大鼠动情周期的改变,Elise检测血清卵泡刺激素(FSH)、雌二醇(E2)水平的改变,流式细胞术检测卵泡颗粒细胞凋亡数量,Western blot检测卵泡颗粒细胞中磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)通路相关蛋白及凋亡相关蛋白的改变。结果:给予补肾养精颗粒干预后,大鼠动情时间较模型组明显缩短(P<0.05),血清FSH水平明显降低(P<0.05),E2水平明显升高(P<0.05),且均有剂量依赖性;同时大鼠卵泡颗粒细胞凋亡数明显减少(P<0.05),且具有剂量依赖性。补肾养精颗粒干预后,磷酸化PI3K及磷酸化Akt蛋白表达上升,抑凋亡基因B淋巴细胞瘤-2(Bcl-2)表达升高,促凋亡Bcl相关x蛋白(Bax)表达降低。结论:补肾养精颗粒通过PI3K/Akt信号通路调节凋亡相关基因,从而达到抑制卵泡颗粒细胞凋亡、改善卵巢功能的目的。

Objective:To investigate the mechanism of Bushen Yangjing Granules regulating follicular granulosa cells apoptosis in model rats with premature ovarian insufficiency(POI). Methods:Rats with POI were modeled by tripterygium glycosides and randomly divided into high,medium and low dosage group of Bushen Yangjing Granules,blank group and non-model group. Different doses of Bushen Yangjing Granules were given for 15 days,and the rats in the blank group and the non-model group were given normal saline.Changes in estrous cycle in rats were continuous observed. Elise was used to detect the changes in serum follicle stimulating hormone(FSH) and estradiol(E2)levels,flow cytometry to detect follicular granulosa cell apoptosis,and Western blot analysis to detect phosphatidylinositol-3-kinase(PI3 K) and protein kinase B(Akt) pathway-associated proteins and apoptosis-related proteins in follicular granulosa cells.Results:After the intervention of Bushen Yangjing Granules,the estrous time of rats was significantly shorter than that of the model group(P< 0.05),serum FSH levels were significantly decreased(P< 0.05),E2 levels were significantly increased(P< 0.05),with dosedependent. At the same time,the number of apoptosis of follicular granulosa cells was significantly reduced(P< 0.05),and it was dose-dependent. After the intervention of Bushen Yangjing Granules,the expression of p-PI3 K and p-Akt protein increased,the expression of B-cell lymphoma-2(Bcl-2) was increased,and the apoptosis-promoting Bcl-related x protein(Bax) expression is reduced. Conclusion:Bushen Yangjing Granules regulated apoptosis-related genes through PI3 K/Akt signaling pathway to achieve the purpose of inhibiting follicular granulosa cell apoptosis and improving ovarian function.