中国比较医学杂志2025，Vol.35Issue(4)：69-78,10.DOI:10.3969/j.issn.1671-7856.2025.04.007
Chrm3通过MAPK/ERK途径调节LPS诱导的Lbp-/-小鼠腹腔巨噬细胞炎症
Chrm3 regulates LPS-induced inflammation in peritoneal macrophages in Lbp-/-mice via the MAPK/ERK signaling pathway
摘要
Abstract
Objective To investigate the role of cholinergic receptor muscarinic 3(Chrm3)in regulating lipopolysaccharide(LPS)-induced inflammation in peritoneal macrophages in lipopolysaccharide binding protein(LBP)-knockout(Lbp-/-)mice.Methods Peritoneal macrophages were isolated from wild-type and Lbp-/-mice to establish an LPS-induced inflammation model.Chrm3 expression in Lbp-/-mouse peritoneal macrophages was inhibited by 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide(4-damp)and small interfering(siRNA)and Chrm3 overexpression was achieved by lentivirus transfection.For 4-damp inhibition,cells were divided into control,LPS,and inhibitor groups,and for siRNA transfection,cells were divided into control,LPS,si-normal control group,and si-Chrm3 groups.For overexpression,cells were divided into control,LPS,negative control,and overexpression groups.Changes in Chrm3 in response to LPS stimulation were verified by Western blot.The effects of 4-damp,si-Chrm3,and lentivirus on cell inflammation and survival were confirmed by Cell Counting Kit-8,quantitative polymerase chain reaction,and Western blot assays.Results Chrm3 protein expression was significantly elevated in Lbp-/-peritoneal macrophages post-LPS stimulation(P＜0.001),whereas there was no notable change in wild-type cells.The cell survival rate was significantly increased in the 4-damp and si-Chrm3 groups(P＜0.05,P＜0.01),and cell survival was significantly reduced in the overexpression group(P＜0.01).Furthermore,4-damp and si-Chrm3 significantly reduced expression levels of the inflammatory factors tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6(P＜0.01,P＜0.001),and phospho-extracellular signal-regulated kinase(p-ERK)(P＜0.01,P＜0.001),which are associated with cell damage and inflammation.In contrast,TNF-α,IL-1β,IL-6(P＜0.001),and p-ERK protein(P＜0.001)were significantly elevated in the overexpression group.Conclusions LPS stimulation upregulated the expression of Chrm3 and proinflammatory cytokines in Lbp-/-peritoneal macrophages.Specific downregulation of Chrm3 by 4-damp and si-Chrm3 significantly decreased LPS-induced proinflammatory cytokines in Lbp-/-peritoneal macrophages,while upregulation of Chrm3 using overexpressing lentivirus significantly elevated the expression of related inflammatory factors.Chrm3 is implicated in the regulation of the LPS-induced inflammation response in peritoneal macrophages in Lbp-/-mice.关键词
毒蕈碱受体/Lbp-/-小鼠/腹腔巨噬细胞/4-damp/干扰RNA/过表达/MAPK信号通路Key words
Chrm3/Lbp-/-mice/peritoneal macrophages/4-damp/RNAi/overexpression/MAPK signaling pathway分类
医药卫生引用本文复制引用
陈志达,付彬,李思迪,刘赛,郭中坤,张悦,王可洲..Chrm3通过MAPK/ERK途径调节LPS诱导的Lbp-/-小鼠腹腔巨噬细胞炎症[J].中国比较医学杂志,2025,35(4):69-78,10.基金项目
山东省医学科学院医药卫生科技创新工程 （）
济南市科技局"高校20条"(2021GXRC011) （2021GXRC011）
山东省生猪产业技术体系(SDAIT-08-17). （SDAIT-08-17）
