温阳健脾方调节AMPK/mTOR/ULK1通路上调RSA小鼠孕期自噬水平改善其蜕膜化的机制研究  

Study on the mechanism of Wenyang Jianpi decoction regulating AMPK/mTOR/ULK1 pathway,up-regulating autophagy level in RSA mouse during pregnancy and improving decidualization

作　　者：赵贵霜 刘娟 李孟瑶 姜丽娟 ZHAO Guishuang;LIU Juan;LI Mengyao;JIANG Lijuan(First Clinical Medical College,Yunnan University of Traditional Chinese Medicine,Kunming 650000,China)

机构地区：[1]云南中医药大学第一临床医学院,云南昆明650000 [2]云南中医药大学第一附属医院伦理委员会,云南昆明650021

出　　处：《陕西中医》2024年第9期1155-1160,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金地区科学基金资助项目(82060882)。

摘　　要：目的:探讨温阳健脾方通过影响AMPK/mTOR/ULK1信号轴调控RSA小鼠孕期自噬水平,从而对其蜕膜化影响的机制研究。方法:CBA/J雌鼠与BALB/c雄鼠合笼建立空白组10只,CBA/J雌鼠与DBA/2雄鼠合笼建立RSA小鼠模型,随机将其分为模型组、温阳健脾方中剂量组,10只/组;于妊娠第1天开始,温阳健脾方组予温阳健脾方13.96 g/(kg·d)灌胃,空白组及模型组予以等体积0.9%氯化钠溶液,连续干预14 d后眼球取血,取蜕膜组织,计算胚胎丢失率,ELISA测定各组小鼠血清中胰岛素样生长因子结合蛋白1(IGFBP-1)和催乳素(PRL),HE染色观察小鼠子宫内膜形态,Western blot法测定小鼠蜕膜组织AMPK、p-AMPK、p-mTOR、p-ULK1、LC3Ⅱ/LC3Ⅰ、Beclin1、p62表达。结果:与空白组相比,RSA模型组胚胎吸收率显著降低(P<0.01);血清中IGFBP-1、PRL降低,LC3Ⅱ/LC3Ⅰ蛋白相对表达量显著下调(P<0.001),p-AMPK/AMPK、Beclin1蛋白相对表达量明显下降,mTOR、ULK1、p62蛋白表达水平显著上调(P<0.01);与模型组比较,温阳健脾方组胚胎吸收率明显降低(P<0.05);血清中IGFBP-1、PRL水平上升(P<0.05),LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK、Beclin1蛋白相对表达量均上调,mTOR、ULK1、p62蛋白相对表达量下调(P<0.05)。结论:温阳健脾方可通过AMPK/mTOR/ULK1通路促进RSA小鼠孕期自噬水平从而改善其蜕膜化,有效提高妊娠率。Objective:To study the influence of Wenyang Jianpi decoction on autophagy level of recurrent spontaneous abortion(RSA)mice during pregnancy by influencing AMPK/mTOR/ULK1 signal axis.Methods:5 CBA/J female rats and BALB/c male rats were caged to establish 10 normal control rats.CBA/J female mice were caged with DBA/2 male mice to establish RSA mouse model,which were randomly divided into model group and middle dose group of Wenyang Jianpi decoction,with 10 mice in each group.From the first day of pregnancy,the Wenyang Jianpi decoction group was given a moderate dose of Wenyang Jianpi decoction 13.96 g/(kg·d),while the control group and the model group were given the same volume of normal saline.After 14 days of continuous intervention,eyeball blood was taken,decidual tissue was taken,the embryo loss rate was calculated,and insulin-like growth factor-1(IGFBP-1),Prolactin(PRL)and HE staining were used to observe the morphology of mouse endometrium,and Western blot was used to determine the expressions of AMPK,p-AMPK,p-mTOR,p-ULK1,LC3Ⅱ/LC3Ⅰ,Beclin1 and p62 in mouse decidua.Results:Compared with the control group,the embryo absorption rate of RSA model group decreased significantly(P<0.01).Serum IGFBP-1 and PRL decreased,the relative expression of LC3Ⅱ/LC3Ⅰprotein decreased significantly(P<0.001),the relative expression of p-AMPK/AMPK and Beclin1 protein decreased significantly,and the expression levels of mTOR,ULK1 and p62 protein increased significantly(P<0.01).Compared with the model group,the embryo absorption rate of Wenyang Jianpi decoction group decreased significantly(P<0.05).The levels of serum IGFBP-1 and PRL increased(P<0.05),while the relative expression levels of LC3Ⅱ/LC3Ⅰ,p-AMPK/AMPK and Beclin1 all increased,while the relative expression levels of mTOR,ULK1 and p62 decreased(P<0.05).Conclusion:Wenyang Jianpi decoction can promote the autophagy level of RSA mice during pregnancy through AMPK/mTOR/ULK1 pathway,thus improving decidualization and effectively increasing the pregnancy rate.

关 键 词：复发性流产 自噬 蜕膜化 温阳健脾方 AMPK/mTOR/ULK1信号通路 阴阳平衡 胰岛素样生长因子结合蛋白1 催乳素 

分 类 号：R714.21[医药卫生—妇产科学]