滑胎安胎协定方促进滋养细胞迁移侵袭治疗复发性流产的作用机制研究  

The mechanism of action of Chinese medicine Huatai Antai decoction in promoting trophoblast migration and invasion in treatment of recurrent spontaneous abortion

作　　者：张旻轶 楼毅云[2] ZHANG Minyi;LOU Yiyun(Department of Gynecology,Tongde Hospital of Zhejiang Province,Hangzhou 310012,China;不详)

机构地区：[1]浙江省立同德医院妇科,杭州310012 [2]杭州市中医院妇二科

出　　处：《浙江医学》2023年第14期1486-1491,I0004,共7页Zhejiang Medical Journal

基　　金：浙江省中医药科技计划青年人才基金项目(2022ZQ021);国家自然科学基金青年科学基金项目(81801475);浙江省中医药科学计划项目(2022ZA111)。

摘　　要：目的探讨滑胎安胎协定方促进滋养细胞迁移侵袭治疗复发性流产(RSA)的作用机制。方法体外培养蜕膜巨噬细胞(DM)并分为中药干预组(加入滑胎安胎协定方含药血清)、IL-4诱导组和空白对照组;采用流式细胞术检测DM分型,ELISA法检测DM中粒细胞集落因子(G-CSF)含量。共培养DM与滋养细胞HTR-8/SVneo细胞,分为DM组、DM+中药组、G-CSF组和空白对照组;采用细胞计数试剂盒-8法检测细胞存活率,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力,Western blot检测磷脂酰肌醇三羟基激酶(PI3K)/蛋白激酶B(AKT)/细胞外调节蛋白激酶(ERK)和上皮间质转化(EMT)相关因子蛋白表达水平。结果DM实验显示,与空白对照组比较,中药干预组、IL-4诱导组DM数量均明显减少(均P<0.05),M2型DM数量均明显增加(均P<0.05),而M1型DM数量比较差异均无统计学意义(均P>0.05);DM中G-CSF含量均明显升高(均P<0.05)。滋养细胞实验显示,与空白对照组比较,DM组、DM+中药组、G-CSF组滋养细胞存活率、迁移率均明显升高(均P<0.05),滋养细胞侵袭数量均明显增多(均P<0.05);DM组、DM+中药组、G-CSF组滋养细胞p-AKT/AKT、p-ERK1/2/ERK1/2、波形蛋白表达水平均明显升高(均P<0.05),DM+中药组、G-CSF组上皮细胞钙黏蛋白表达水平均明显降低(均P<0.05)。结论滑胎安胎协定方可通过促进DM分泌G-CSF来激活PI3K/AKT/ERK信号通路和EMT进程,进而调控滋养细胞增殖、迁移和侵袭,从而防止RSA的发生。Objective To explore the mechanism of action of Chinese medicine Huatai Antai decoction in promoting trophoblast migration and invasion in the treatment of recurrent spontaneous abortion(RSA).Methods Human decidual macrophage(DM)were cultured in vitro and divided into three groups:the Chinese medicine intervention group(adding drugcontaining serum from Huatai Antai decoction),the interleukin 4(IL-4)induction group,and the blank control group.DM typing was detected by flow cytometry,and the content of granulocyte colony-stimulating factor(G-CSF)in cells was detected by ELISA.DMs and human trophoblast HTR-8/SVneo cells were co-cultured and divided into DM group,DM+Chinese medicine group,G-CSF group and blank control group.Cell counting kit-8 method was used to detect cell activity,scratch test was used to detect cell migration,Transwell test was used to detect cell invasion,and Western blot was used to detect the protein expression levels of phosphoinositide-3 kinase/protein kinase B/extracellularly regulated protein kinases(PI3K/AKT/ERK)and epithelial-mesenchymal transition(EMT)-related factors.Results Compared with the blank control group,the number of DM in the Chinese medicine intervention group and the IL-4 induced group were all significantly lower(all P<0.01),and the number of M2-type DM were all significantly increased(all P<0.05),whereas there was no significant difference in the number of M1-type DM(P>0.05),and the content of G-CSF in the DM was significantly high(all P<0.05).When DM were cocultured with trophoblast HTR-8/SVneo cells,compared with the blank control group,trophoblast survival and migration rates in the DM group,DM+Chinese medicine group,and G-CSF group were all significantly higher(all P<0.05),and the number of trophoblast invasion was all significantly increased(all P<0.05);the expression levels of p-AKT/AKT,p-ERK1/2/ERK1/2 and vimentin in trophoblast were all significantly higher(all P<0.05),and expression levels of epithelial cell calmodulin were all significantly lower in the DM+Chines

关 键 词：滑胎安胎协定方 复发性流产 蜕膜巨噬细胞 滋养细胞 粒细胞集落因子 

分 类 号：R285[医药卫生—中药学]