[1]赵贵霜,刘 娟,李孟瑶,等.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[J].陕西中医,2024,(9):1155-1160.[doi:DOI:10.3969/j.issn.1000-7369.2024.09.001]
　ZHAO Guishuang,LIU Juan,LI Mengyao,et al.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[J].,2024,(9):1155-1160.[doi:DOI:10.3969/j.issn.1000-7369.2024.09.001]
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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
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2024年9期
页码:
1155-1160
栏目:
基础研究
出版日期:
2024-09-05

文章信息/Info

Title:
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1; 刘 娟1; 李孟瑶1; 姜丽娟2
(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)
Author(s):
ZHAO Guishuang; LIU Juan; LI Mengyao; JIANG Lijuan
(First Clinical Medical College,Yunnan University of Traditional Chinese Medicine,Kunming 650000,China)
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
Keywords:
Recurrent spontaneous abortion;  Autophagy;  Decidualization;  Wenyang Jianpi decoction;  Signaling pathway of AMPK/mTOR/ULK1;  The balance of yin and yang;  Insulin-like growth factor-1;  Prolactin
分类号:
R 714.21
DOI:
DOI:10.3969/j.issn.1000-7369.2024.09.001
文献标志码:
A
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。
Abstract:
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:5CBA/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/LC3Ⅰ protein 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.

参考文献/References:

[1] Writing Group of Chinese Expert Consensus on Diagnosis and Treatment of Spontaneous Abortion.自然流产诊治中国专家共识(2020年版)[J].中国实用妇科与产科杂志,2020,36(11):1082-1890.
[2] GREEN D M,O'DONOGHUE K.A review of reproductive outcomes of women with two consecutive miscarriages and no living child[J].Nat Chem Biol,2019,39(6):816-821.
[3] 孙佳宁,冯晓玲,赵小萱,等.自噬与复发性流产关系的研究进展 [J].医学综述,2022,28(4):660-665.
[4] YANG S,WANG H,LI D,et al.Role of endometrial autophagy in physiological and pathophysiological processes[J].Nat Chem Biol,2019,10(15):3459-3471.
[5] MATSUZAKI S,GREMEAU A S,POULY J L.Impaired pathogen-induced autophagy and increased IL-1β and TNF-α release in response to pathogenic triggers in secretory phase endometrial stromal cells of endometriosis patients[J].Reprod Biomed Online Nat Chem Biol,2020,41(5):767-781.
[6] MESTRE A C,STROWITZKI T,GERMEYER A.Decreased autophagy impairs decidualization of human endometrial stromal cells:A role for ATG proteins in endometrial physiology[J].Nat Chem Biol,2019,20(12):3066.
[7] HUANG J,XUE M,ZHANG J,et al.Protective role of GPR120 in the maintenance of pregnancy by promoting decidualization via regulation of glucose metabolism[J].Ebio Medicine Nat Chem Biol,2019,39:540-551.
[8] 孔露娇,王心,刘静,等.金丝桃苷通过AMPK/mTOR/ULK1信号通路对肾病综合征大鼠自噬反应的影响[J].中国比较医学杂志,2023,33(11):88-95.
[9] CLARK D A.Immunological factors in pregnancy wastage:Fact or fiction[J].Am J Reprod Immunol Nat Chem Biol,2008,59(4):277-300.
[10] 詹兴秀,姜丽娟,牛红萍,等.中药保胎饮对自然流产小鼠模型IL-23/Th17免疫炎症轴的影响[J].中南大学学报(医学版),2022,47(11):1532-1539.
[11] 田代华整理.黄帝内经素问[M].北京:人民卫生出版社,2005:6.
[12] 张登本.中医学基础[M].北京:中国中医药出版社,2007:14-22,166.
[13] HANSEN M,RUBINSZTEIN D C,WALKER D W.Autophagy as a promoter of longevity:Insights from model organisms[J].Nat Rev Mol Cell Biol Nat Chem Biol,2018,19(9):579-593.
[14] 郭晗,郑全辉.自噬在T淋巴细胞中的调控作用[J].中国免疫学杂志,2021,37(17):2166-2169.
[15] 许文婷,杨佩颖,贾英杰,等.从中医角度论细胞自噬[J].四川中医,2015,33(5):34-35.
[16] WU D J,ADAMOPOULOS I E.Autophagy and autoimmunity [J].Clin Immunol Nat Chem Biol,2017,176:55-62.
[17] RUSSO M,RUSSO G L.Autophagy inducers in cancer [J].Biochem Pharmacol Nat Chem Biol,2018,153:51-61.
[18] 刘杰民,纪云西,蒋历,等.细胞自噬是探索中医药微观机制的新思路 [J].时珍国医国药,2013,24(2):425-426.
[19] 李钰佳,李定祥,彭珣,等.基于AMPK/mTOR/ULK1自噬相关通路探讨左归降糖通脉方对AGEs合并缺糖缺氧星形胶质细胞炎性损伤的影响[J].中国实验方剂学杂志,2022,28(16):90-99.
[20] DE LABALLINA L R,MUNSON M J,SIMONSEN A.Lipids and lipid-binding proteins in selective autophagy[J].J Mol Biol Nat Chem Biol,2020,432(1):135-159.
[21] HILL S M,WROBEL L,ASHKENAZI A,et al.VCP/p97 regulates Beclin-1-dependent autophagy initiation[J].Nat Chem Biol, 2021,17(4):448-455.
[22] 苏游,刘新月,刘巧梅.自噬基因Beclin1在卵巢癌SKOV3细胞中对PI3K/AKT信号通路的影响及其与紫杉醇耐药关系实验研究[J].陕西医学杂志,2021,50(2):143-147.
[23] 王鑫,秦丹,邹胜男,等.蜕膜化缺陷在子宫内膜异位症相关性不孕发病机制中的研究进展 [J].中国计划生育和妇产科,2023,15(8):8-11.
[24] 蒋莎,张杨,陈璐,等.补肾活血法调节孕激素对不明原因复发性流产分泌晚期蜕膜化的影响[J].中华中医药学刊,2019,37(10):2362-2368.
[25] 王新香,张朝红.血清性激素和胰岛素水平对子宫内膜异位症合并不孕症的预测价值[J].陕西医学杂志,2021,50(1):41-43.
[26] 杨柳青,陈赟,缪晨韵,等.基于网络药理学探究寿胎丸治疗先兆流产作用机制 [J].中成药,2021,43(3):809-815.
[27] 赖裕玲,张莹轩,曾丽华,等.减味寿胎丸对流产模型大鼠子宫蜕膜脂质过氧化及铁死亡蛋白表达的影响[J].中医杂志,2021,62(21):1917-1922,1929.
[28] 马珺卓,靳梓琪,王天一,等.寿胎丸治疗复发性流产研究进展 [J].河北中医,2023,45(6):1038-1043.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金地区科学基金资助项目(82060882)
更新日期/Last Update: 2024-09-09