摘要
目的探究二甲双胍(metformin,MET)与miR-129-5p在脓毒症相关肺损伤中的相互作用及其可能的作用机制。方法脂多糖(lipopolysaccharide,LPS)诱导MHS细胞构建脓毒症细胞模型,盲肠结扎穿孔建立脓毒症动物模型,MET干预细胞、动物模型,CCK-8、流式细胞术检测细胞活力及凋亡,RT-qPCR测定miR-129-5p表达变化,酶ELISA测定TNF-α、IL-6、IL-1β炎症因子表达,HE染色探索肺组织病理学改变,并测量肺干湿比重(D/W);Western blot测定PI3K、p-PI3K、AKT、p-AKT蛋白水平。结果与模型组相比,MET处理可抑制细胞凋亡并提高细胞活力。还可降低TNF-α、IL-6、IL-1β,上调miR-129-5p的表达。而过表达miR-129-5p能改善LPS诱导的炎症反应,敲低miR-129-5p则显示出相反的炎症调控反应,并减弱MET的保护作用。在小鼠模型中,MET处理可下调TNF-α、IL-6、IL-1β水平,减轻肺水肿,提高其存活率。此外,MET可使LPS诱导的PI3K/AKT通路激活,上调p-PI3K、p-AKT蛋白的表达。结论MET可能通过调控miR-129-5p水平,激活PI3K/AKT通路,减轻脓毒症相关肺损伤炎症反应,降低细胞凋亡,起到肺保护作用。
Aim To explore the interaction between metformin(MET)and miR-129-5p in sepsis lung injury and its possible mechanism.Methods MHS cells were induced by lipopolysaccharide(LPS)to establish the sepsis cell model,and animal model was established by cecal ligation and puncture(CLP),both cell model and animal model were intervened by MET.Cell viability and apoptosis were detected by CCK-8 and flow cytometry,the expression of miR-129-5p was determined by fluorescence quantitative PCR(RT qPCR),and the expressions of IL-1β,IL-6,TNF-αwere determined by enzyme-linked immunosorbent assay(ELISA).The pathological changes of lung tissue,and the specific gravity of lung dry and wet(D/W)were measured using Hematoxylin eosin staining.The expression of PI3K,p-PI3K,AKT,p-AKT was determined by Western blot.Results Compared with the model group,MET treatment could inhibit cell apoptosis and improve cell viability.It can also reduce the pro-inflammatory factor(IL-1βIL-6 TNF-α)and up-regulate the expression of miR-129-5p.While overexpression of miR-129-5p could improve LPS induced inflammatory response,knockdown of miR-129-5p showed the opposite inflammatory regulatory response and attenuated the protective effect of MET.In a mouse model,met treatment down-regulated IL-1β,IL-6,TNF-αto reduce pulmonary edema and improve its survival rate.In addition,MET could activate the PI3K/AKT pathway induced by LPS and upregulate the expression of p-PI3K and p-AKT protein.Conclusions MET may play a role in lung protection by regulating the level of miR-129-5p,activating PI3K/AKT pathway,alleviating the inflammatory response of sepsis lung injury and reducing the apoptosis of MHS cells.
作者
袁媛
刘金虹
袁江汉
周发春
YUAN Yuan;LIU Jin-hong;YUAN Jiang-han;ZHOU Fa-chun(Critical Care Dept,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China;Dept of Pharmacy,Tianjin Baodi Hospital,Baodi Clinical College of Tianjin Medical University,Tianjin 301800,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2023年第11期2089-2095,共7页
Chinese Pharmacological Bulletin
基金
重庆市自然科学基金面上项目(No cstc2020jcyj-msxmX1096)。
