左归丸对6.0 Gyγ射线损伤大鼠卵巢组织凋亡和p53蛋白表达的影响

Effects of Zuogui pill on ovarian apoptosis and p53 protein expression in rats damaged by 6.0 Gyγradiation

目的探讨6.0 Gyγ射线对性成熟期大鼠血清性激素水平、卵巢组织凋亡的影响,以及左归丸的干预效应。方法将60只8周龄雌性SD大鼠,随机分成正常对照组、辐射组、补佳乐组、左归丸高剂量组、左归丸低剂量组和左归丸高剂量+补佳乐组,每组10只。γ射线一次性照射(6.0 Gy,LD40)24小时后给予补佳乐、左归丸高剂量、低剂量、左归丸高剂量+补佳乐灌胃,每日一次,连续用药21天后,用ELISA法检测治疗前后血清激素(卵泡雌激素、黄体生成素和雌二醇);苏木精—伊红观察卵巢的组织形态变化;流式细胞仪检测卵巢组织凋亡率以及免疫组化检测凋亡蛋白p53表达。结果与正常组比较,辐射组大鼠卵泡卵母细胞和颗粒细胞出现核固锁,初级卵泡增多,窦状卵泡比例下降(P<0.01),血清卵泡雌激素水平升高(P<0.05),卵巢组织凋亡率明显上升(P<0.01),卵巢组织p53蛋白表达明显增加(P<0.01);与模型组比较,各治疗组窦状卵泡比例增加(P<0.01),初级卵泡增多,血清卵泡雌激素水平下降(P<0.05),卵巢组织凋亡率减少(P<0.01),卵巢组织p53蛋白表达明显减少(P<0.05),尤其是左归丸高剂量组+补佳乐作用显著(P<0.01,P<0.05)。结论左归丸可通过抑制卵巢组织凋亡和p53蛋白表达,促进初级卵泡发育,增加血清雌二醇含量,降低血清卵泡雌激素水平,达到缓解辐射损伤卵巢的作用。

Objective To investigate the effect of 6.0 Gyγeffects of radiation on serum sex hormone levels and ovarian apoptosis in rats during sexual maturity and intervention effect of Zuogui pill.Methods All the eight weeks old female SD rats were randomly divided into the normal group,the radiation group,the Bujiale group,the Zuogui pill high dose group,the Zuogui pill low dose group and the Zuogui pill high dose+Bujiale group,with 10 rats in each group.γafter one-time irradiation(6.0gy,LD40)for 24h,the Bujiale,Zuogui pill high-dose,low-dose,Zuogui pill high-dose+Bujiale groups were given by gavage once a day for 21 days.Serum hormones(FSH,LH and E 2)were detected by ELISA before and after treatment,and histological changes of ovary were observed by hematoxylin eosin(HE).The apoptosis rate of ovarian tissue was detected by flow cytometry and the expression of p53 protein was detected by immunohistochemistry.Results Compared with the normal group,the oocytes and granulosa cells in the radiation model group showed that the nuclear lock and the number of primary follicles were increased,the proportion of antral follicles was decreased(P<0.01),the serum FSH level was increased(P<0.05),the apoptosis rate of ovarian tissue increased significantly(P<0.01),and the expression of p53 protein in ovarian tissue increased significantly(P<0.01);Compared with the model group,the proportion of antral follicles was increased(P<0.01),the number of primary follicles was increased,the serum FSH level was decreased(P<0.05),the apoptosis rate of ovarian tissue was decreased(P<0.01),and the expression of p53 protein in ovarian tissue was decreased significantly(P<0.05),especially in the Zuogui pill high-dose+Bujiale(P<0.01,0.05).Conclusion Zuogui pill can reduce ovarian apoptosis and p53 protein expression induced by radiation,increase E 2 content,reduce FSH,promote primary follicle development,and alleviate ovarian injury.