摘要
目的探讨骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)对细菌脂多糖(LPS)诱导流产的小鼠机体炎症反应的影响及保胎作用。方法选取清洁级SD孕鼠构建Control组(PBS处理)、LPS组(LPS诱导处理)、LPS+BMMSCs组,观察小鼠胚胎发育情况。在孕13天对各组小鼠眼球取血后处死孕鼠,取蜕膜组织;ELISA检测血清内炎性因子(TNF-α、MCP-1、IL-10、IL-17A)水平;确定各组小鼠的流产率和胚胎吸收率;苏木精-伊红染色(HE)方法观察各组小鼠蜕膜组织病理学变化;流式细胞仪检测各组小鼠蜕膜组织内T细胞亚群CD8^(+)、CD14^(+)、CD40^(+)百分比;Western-blot检测蜕膜组织内NF-κB/NLRP3信号通路关键蛋白NF-κB、NLRP3、Caspase-1蛋白表达。构建LPS组、Neferine组(NF-κB/NLRP3信号通路抑制剂)、LPS+BMMSCs+Neferine组,Western-blot检测各组小鼠蜕膜组织内NF-κB、NLRP3、Caspase-1表达。结果LPS组孕鼠全部发生流产现象,胎盘结构出现损坏;BMMSCs干预后流产现象得到改善;与Control组相比,LPS组孕鼠流产率、胚胎吸收率均高达100.00%(P<0.05),血清内炎性因子(TNF-α、MCP-1、IL-17A)水平、细胞因子(CD8^(+)、CD14^(+)、CD40^(+))水平、Markin评分、NF-κB、NLRP3、Caspase-1表达均明显提升,IL-10水平下降(P<0.05);BMMSCs干预可以降低经LPS诱导孕鼠的流产率以及胚胎吸收率(P<0.05),降低经LPS诱导孕鼠血清炎性因子(TNF-α、MCP-1、IL-17A)、细胞因子(CD8^(+)、CD14^(+)、CD40^(+))水平、Markin评分及NF-κB、NLRP3、Caspase-1表达,提升IL-10水平(P<0.05)。NF-κB/NLRP3信号通路被阻断后NF-κB、NLRP3、Caspase-1表达降低(P<0.05);BMMSCs干预可以进一步降低NF-κB、NLRP3、Caspase-1表达(P<0.05)。结论移植间充质干细胞可能通过抑制NF-κB/NLRP3信号通路活性来减少巨噬细胞、T细胞活化以及炎症因子的释放,减轻蜕膜组织病理损伤,从而缓解脂多糖诱导的小鼠流产现象,起到保胎作用。
Objective To investigate the effect of bone marrow mesenchymal stem ells(BMMSCs)on the inflammatory response and fetal protection of bacterial lipopolysaccharide(LPS)induced abortion mice.Methods Clean grade SD pregnant mice were selected to construct a control group(PBS treatment),LPS group(LPS induction treatment),and LPS+BMMSCs group;Observed the development status of mice embryos.On the 13th day of pregnancy,blood was collected from the eyeballs of each group of mice,and the pregnant mice were euthanized and decidual tissue was taken;ELISA detection of serum inflammatory factors(TNF-α,MCP-1,IL-10,IL-17A)levels;observed the abortion rate and embryo absorption rate of each group of mice;Hematoxylin eosin staining(HE)was used to observed the changes of decidual histopathology in each group;flow cytometry was used to detect the percentage of T cell subpopulations CD8^(+),CD14^(+),and CD40^(+)in the decidual tissue of mice in each group;Western-blot detection of NF-κB/NLRP3 signaling pathway key protein NF-κB,NLRP3 and Caspase-1 protein expression.Constructed LPS group,Neferine group(NF-κB/NLRP3 signaling pathway inhibitor)and LPS+BMMSCs+Neferine group;Western-blot detection of in decidual tissue of mice in each group NF-κB,NLRP3 and Caspase-1 expression.Results All pregnant mice in the LPS group had the abortion phenomenon,with substantial damage to the placental structure;after intervention with BMMSCs,the phenomenon of miscarriage in pregnant mice were improved;compared with the control group,the miscarriage rate and embryo absorption rate in the LPS group were both as high as 100.00%(P<0.05),and serum inflammatory factors(TNF-α、MCP-1,IL-17A)levels,cytokine(CD8^(+),CD14^(+),CD40^(+))levels,Markin score,the expression of NF-κB,NLRP3 and Caspase-1 was significantly increased,while the level of IL-10 decreased(P<0.05);BMMSCs intervention can significantly reduce the abortion rate and embryo absorption rate of LPS induced pregnant mice(P<0.05),and reduce the serum inflammatory factors(TNF-α,MCP-1,IL-17A),
作者
宋艳引
王觅
Song Yanyin;Wang Mi(Department of Obstetrics and Gynecology,the Eco-city Hospital of Tianjin Fifth Central Hospital,Tianjin 300480;Department of Obstetrics and Gynecology,Hebei General Hospiual,Shijiazhuang Hebei 050000,P.R.China)
出处
《中国计划生育和妇产科》
2024年第2期60-65,F0003,共7页
Chinese Journal of Family Planning & Gynecotokology
基金
2018年度河北省医学科学研究重点课题计划(项目编号:20180053)。
