摘要
目的探讨毓麟珠调控TSC/Akt通路改善氧化应激,进而改善卵泡发育的机制。方法动情周期正常的84只雌性BALB/c小鼠随机分为空白组、模型组、雌激素组、毓麟珠组(低、中、高剂量组),每组14只。除空白组外,其余五组建立免疫性早发性卵巢功能不全(primary ovarian insufficiency,POI)模型。造模成功后,除空白组、模型组外,其余各组连续6周给予相应药物灌胃治疗。治疗后,观察卵巢组织病理状态,检测卵巢组织的超氧化物歧化酶(superoxide dismutase,SOD)、活性氧(reactive oxygen species,ROS)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)和丙二醛(malondialdehyde,MDA)的水平,蛋白免疫印迹法检测卵巢组织中RheB、P-s6k1、P-4E-BP1蛋白含量的表达,免疫组化检测卵巢组织中TSC1、TSC2、Akt的蛋白表达。结果造模成功后,模型组小鼠卵巢、子宫质量降低;给药后,毓麟珠高剂量组小鼠左右卵巢以及子宫质量均增加(P<0.01)。与模型组相比,各给药组卵泡数量明显增加,卵巢组织形态明显改善。与空白组相比,模型组氧化因子表达水平较高,抗氧化因子表达水平明显较低(P<0.05);与模型组相比,各给药组ROS、MDA明显升高,GSH-Px、SOD明显降低(P<0.05)。与空白组相比,模型组Akt蛋白表达降低,TSC1、TSC2表达升高;与模型组相比,各给药组Akt蛋白表达明显升高,TSC1、TSC2蛋白表达明显降低。与空白组相比,模型组RheB、P-s6k1、P-4E-BP1表达降低(P<0.05);与模型组相比,各给药组RheB、P-s6k1、P-4E-BP1蛋白表达升高,以P-4E-BP1升高最明显(P<0.01)。结论毓麟珠通过调控TSC/Akt信号通路抑制微环境氧化应激,这可能是其改善卵母细胞质量,促进卵母细胞成熟的作用机制。
Objective The destination of this paper was to discuss the regulation of TSC/Akt pathway by Yulinzhu to improve oxidative stress,and then improve the mechanism of follicle development.Methods 84 female BALB/c mice with normal emotional cycle were selected,they were randomly divided into blank group,model group,estrogen group and Yulinzhu group(low,medium and high dose group).Except for the blank group,five other groups established immune POI models.After the model established,except for the blank group,the model group,the other groups were given corresponding drugs for 6 weeks.Observe the pathological state of ovarian tissue by HE staining,observe the oxidative stress state of ovary by detecting the levels of superoxide dismutase(SOD),active oxygen(ROS),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)in ovarian tissues,and detect the expression of RheB,P-s6k1,P-4E-BP1 protein content in ovarian tissues by Western blot,TSC1,TSC2,Akt protein expression in ovarian tissues was detected by immunohistochemistry.Results The weight of ovary and uterus in mice showed that the quality of ovary in the model group was decreased,after administration.The quality of left and right ovaries and uterus of mice in the Yulinzhu high dose group increased,difference was statistically significant(P<0.01).The HE results show that compared with the model group,The number of follicles in each group increased significantly,the morphology of ovarian tissue was obviously improved;the results of oxidative stress factor test showed that the expression of oxidative stress factor was higher in the model group,the expression of antioxidant factors was significantly lower,after administration,the ROS,MDA increased significantly,GSH-Px,SOD significantly reduced,difference was statistically significant(P<0.05);the immunohistochemical results showed that Akt protein expression was decreased and TSC1,TSC2 protein expression were increased in the model group.After administration,the Akt protein expression was increased significantly,the TSC1,TSC2 protein expression were decreased significantly;the Western blotting showed that the expressions of RheB,P-s6k1,P-4E-BP1 in the model group were decreased,the expressions of RheB,P-s6k1,P-4E-BP1 protein in the administration group were increased significantly,difference was statistically significant(P<0.05).Conclusion Yulinzhu regulates the TSC/Akt signal pathway,through the mechanism of inhibiting microenvironment oxidative stress.This may be the mechanism for improving oocyte quality and promoting oocyte maturation.
作者
魏茂林
杨珍
董晓英
冯逊逊
WEI Maolin;YANG Zhen;DONG Xiaoying;FENG Xunxun(School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China)
出处
《环球中医药》
CAS
2021年第12期2127-2134,共8页
Global Traditional Chinese Medicine
基金
北京市自然科学基金面上项目(7212162)
北京中医药薪火传承“3+3”工程(2012-SZ-C-41)。
