基于PI3K/Akt/mTOR通路探讨白藜芦醇改善大鼠化疗性卵巢损伤的机制

Mechanism of Resveratrol on Improving Chemotherapy-induced Ovarian Injury in Rats through PI 3K/Akt/mTOR Signaling Pathway

目的基于PI 3K/Akt/mTOR信号通路探讨白藜芦醇改善大鼠的化疗性卵巢损伤的分子机制。方法30只雌性Wistar大鼠随机分为对照组、化疗组(顺铂组)、化疗给药组(白藜芦醇+顺铂组),实验时间共21天。化疗给药组大鼠给予白藜芦醇25mg/(kg·d)灌胃,化疗组给予0.9%NaCl溶液。在实验的第15天,除对照组,其余两组均给予单剂量5mg/kg的顺铂腹腔注射。1周后取大鼠卵巢制作石蜡切片,采用苏木精-伊红染色(HE)观察卵巢形态及卵泡发育情况。原位缺口末端标记法(TUNEL)检测分析卵巢组织的调亡情况。蛋白免疫印迹实验(Western blot)法分析比较卵巢组织中磷脂酰肌醇3-激酶(PI 3K)、蛋白激酶(Akt)、哺乳动物雷帕霉素靶蛋白(mTOR)及其磷酸化水平的蛋白表达差异。结果与对照组比较,化疗组卵巢萎缩,黄体退化,各级生长卵泡数明显减少;卵巢细胞的凋亡数目增多,凋亡指数升高(P<0.01);p-PI 3K/PI 3K、p-Akt/Akt、p-mTOR/mTOR蛋白表达均下降(P<0.01)。化疗给药组与化疗组比较,可见卵巢结构较化疗组有明显改善,间质纤维化程度有所减轻,且可见一定数量的黄体,各级生长卵泡数增多;卵巢细胞凋亡数目减少,凋亡指数降低(P<0.01);p-PI 3K/PI 3K、p-Akt/Akt、p-mTOR/mTOR蛋白表达均上升(P<0.01)。结论白藜芦醇能改善顺铂化疗导致的大鼠卵巢损伤,其机制可能与激活PI 3K/Akt/mTOR信号通路有关。

Objective To investigate the molecular mechanism through PI 3K/Akt/mTOR signaling pathway of resveratrol in improving chemotherapy-induced ovarian function injury in rats.Methods Thirty female Wistar rats were randomly divided into three groups including control,chemotherapy and resveratrol plus chemotherapy groups(n=10 for each group).Rats in the resveratrol plus chemotherapy group were administrated with resveratrol 25mg/(kg·d)by gavage,and rats were given with saline in the chemotherapy group over three weeks.All groups except the control group were treated with 5mg/kg cisplatin in the abdominal cavity on the 15th day.All rats underwent oophorectomy one week after injection of cisplatin.Ovarian morphology and follicles were counted microscopically.The apoptosis of ovarian tissue were detected by Tunel Kit.The expression of PI 3K,p-PI 3K,Akt,p-Akt,mTOR and p-mTOR proteins were determined by Western blot.Results Compared with the control group,ovary atrophied,corpus luteum degenerated,growth follicular number was reduced,apoptotic index was remarkably increased(P<0.01)in the chemotherapy group.The expression of p-PI 3K/PI 3K,p-Akt/Akt and p-mTOR/mTOR protein were significantly decreased(P<0.01).Compared with the chemotherapy group,the structure of ovary was improved obviously in the Resveratrol plus chemotherapy group,apoptotic index was considerably decreased(P<0.01).The expression of p-PI 3K/PI 3K,p-Akt/Akt and p-mTOR/mTOR protein were greatly increased(P<0.01).Conclusion Resveratrol can prevent cisplatin chemotherapy induced ovarian damage in rats,and its mechanism might be related to the regulation of PI 3K/Akt/mTOR signaling pathway.