马鞭草苷通过激活GPR18受体对脓毒症诱导的急性肺损伤的保护作用及机制探究  

Protective effect and mechanism of verbenalin on sepsis-induced acute lung injury by activating GPR18 receptor

作　　者：于鑫 董铮 杨磊[3] 杨文杰[4] YU Xin;DONG Zheng;YANG Lei;YANG Wenjie(Graduate School of Tianjin University of Traditional Chinese Medicine,Department of Critical Care Medicine,Characteristic Medical Center of Chinese People's Armed Police Force,Tianjin 300162,China;不详)

机构地区：[1]天津中医药大学研究生学院,中国人民武装警察部队特色医学中心综合重症医学科,天津300162 [2]中国人民武装警察部队天津市总队医院内二科,天津300162 [3]天津市南开医院急腹症研究所,天津300100 [4]天津市第一中心医院感染科,天津300192

出　　处：《中国急救复苏与灾害医学杂志》2023年第8期1051-1055,共5页China Journal of Emergency Resuscitation and Disaster Medicine

基　　金：天津市自然科学基金青年项目(编号:18JCQN)。

摘　　要：目的 探讨马鞭草苷(Verbenalin)是否对脓毒症诱导的急性肺损伤具有保护作用及潜在机制。方法 实验动物采用C57BL/6小鼠,应用体内盲肠结扎穿孔(CLP)手术建立脓毒症急性肺损伤模型,将30只小鼠随机分为三组,分别为:对照组(Cont组)、模型组(CLP组)和马鞭草苷治疗组(CLP+Verbenalin组);体外采用脂多糖(LPS)刺激小鼠单核细胞RAW264.7细胞构建巨噬细胞M1极化模型;采用HE染色法检测肺组织的病理变化;采用流式细胞仪检测肺组织中巨噬细胞的M1极化;采用酶联免疫吸附测定(ELISA)细胞上清及外周血中IL-1β、TNF-α和IL-6的含量;采用蛋白印迹法(Western blotting)法检测目的蛋白的表达。结果 在体内,与对照组比较,模型组出现肺泡壁增厚、炎症细胞浸润数目明显增加的改变;同时肺组织中M1型巨噬细胞的比例增加;与模型组相比,马鞭草苷可以显著改善肺组织病理损伤和外周血中IL-1β、TNF-α和IL-6的分泌及抑制巨噬细胞的M1极化。在体外,LPS刺激诱导IL-1β、TNF-α和IL-6的分泌增加,以及促进NF-κB信号通路活化;马鞭草苷干预后,炎症因子的分泌和P-NF-κB的磷酸化水平均显著降低;同时,给予O1918预处理抑制G蛋白偶联受体18(GRP18)后,马鞭草苷的抑制M1极化的作用减弱。结论 马鞭草苷可以通过激活GPR18受体抑制NF-κB信号通路的活化,进而降低巨噬细胞M1极化和减轻脓毒症诱导的肺损伤。Objective To investigate whether verbenalin has protective effect on sepsis-induced acute lung injury and its potential molecular mechanism.Methods The acute lung injury model of sepsis was established by cecal ligation and perforation(CLP)in vivo.Thirty C57BL/6 mice were randomLy divided into control group(Cont group),model group(CLP group)and Verbenalin treatment group(CLP+Verbenalin group).In vitro,mice monocyte RAW264.7 cells were stimulated by lipopolysaccharide(LPS)to construct a macrophage M1 polarization model.HE staining was used to detect the pathological changes of lung tissue.M1 polarization of macrophages in lung tissue was detected by flow cytometry.Determination of IL-1β,TNF-α,and IL-6 in cell supernatant and peripheral blood by enzyme linked immunosorbent assay(ELISA).Western blotting was used to detect the expression of the target protein.Results Compared with the control group in vivo,the number of inflammatory cells infiltration in the lung tissues of the model group increased significantly and the proportion of M1 macrophages in lung tissue increased significantly.Compared with the model group,verbenalin can significantly improve the pathological damage of lung tissues and IL-1β,TNF-α,and IL-6 secretion in peripheral blood,and inhibit M1 polarization of macrophages in lung tissues.In vitro,LPS stimulation induces IL-1β,TNF-α,and IL-6,and promotes NF-κB signal pathway activation.Verbenalin can reduce the secretion of inflammatory factors and the phosphorylation level of NF-κB.Meanwhile,after O1918 pretreatment inhibited G protein coupled receptor 18(GRP18),the inhibitory effect of verbenalin on M1 polarization was weakened.Conclusion Verbenalin can inhibit the activation of NF-κB signal pathway vie activating GPR18 receptor,thereby reducing M1 polarization of macrophages and alleviating sepsis-induced acute lung injury.

关 键 词：脓毒症 马鞭草苷 炎症小体 急性肺损伤 

分 类 号：R631[医药卫生—外科学]