脓毒症急性肺损伤模型中肺组织细胞焦亡与大麻素2型受体的相关性分析  

Correlation analysis between the expression of cannabinoid 2 receptor and the related indicators of pyroptosis in sepsis

作　　者：张彬 杨宗斌 李祯 刘安鹏 杨柳[1] 郑锋[1] 郑菲[1] 詹佳[1] Zhang Bin;Yang Zongbin;Li Zhen;Liu Anpeng;Yang Liu;Zheng Feng;Zheng Fei;Zhan Jia(Department of Anesthesiology,Zhongnan Hospital,Wuhan University,Wuhan 430071,China;Department of Anesthesiology,The Lichuan Ethnic Hospital of Traditional Chinese Medicine,Wuhan 445400,China)

机构地区：[1]武汉大学中南医院麻醉科,武汉430071 [2]湖北省利川市民族中医院麻醉科,利川445400

出　　处：《中华急诊医学杂志》2022年第2期185-190,共6页Chinese Journal of Emergency Medicine

基　　金：湖北省自然科学基金(2019CFB690)。

摘　　要：目的探讨在小鼠脓毒症急性肺损伤模型中,焦亡相关指标与大麻素2型受体(cannabinoid 2 receptor,CB2R)表达量的相关性。方法采用随机数字表法把实验小鼠随机分为4组(n=6):假手术组(sham)、轻度脓毒症组(ALIMi)、中度脓毒症组(ALIMo)、重度脓毒症组(ALIS)。采用盲肠结扎穿孔法(cecal ligation and puncture,CLP)建立小鼠的脓毒症急性肺损伤模型。术后12 h采集肺组织标本,测定肺组织湿干重比及肺组织损伤评分;检测肺组织CB2RmRNA及蛋白表达水平、焦亡相关指标(NLRP3、caspasse-1、caspase-11、GSDMD)的mRNA转录水平及肺组织炎症因子(IL-6、TNF-α)的水平;利用SPSS软件进行数据分析,采用单因素方差分析法进行各项指标的多组间比较,分别采用LSD或Games-Howell检验进行两组间比较,利用Pearson法分别分析炎症因子和焦亡相关指标与CB2R之间的相关性。结果通过单因素方差分析获得统计量F值,并进行两组间比较。与sham组比较,在脓毒症模型中,IL-6、TNF-α、CB2RmRNA、CB2R蛋白以及NLRP3、caspase-1、caspase-11、GSDMD mRNA表达升高(P<0.05);与ALIMi组比较,ALIMo组小鼠肺组织IL-6[(277.31±41.07)vs.(140.09±27.56),P<0.05]、TNF-α[(501.09±73.91)vs.(261.36±40.73),P<0.05]、CB2RmRNA[(2.99±0.28)vs.(2.02±0.19),P<0.05]、CB2R蛋白[(0.44±0.08)vs.(0.23±0.05),P<0.05]以及NLRP3[(2.53±0.26)vs.(1.61±0.15),P<0.05]、caspase-1[(6.02±0.35)vs.(3.60±0.38),P<0.05]、caspase-11[(11.43±0.83)vs.(6.30±0.65),P<0.05]、GSDMD[(10.46±0.62)vs.(5.67±0.54),P<0.05]mRNA表达升高;与ALIMo组比较,ALIS组小鼠肺组织中IL-6[(475.90±67.65)vs.(277.31±41.07),P<0.05]、TNF-α[(713.93±58.85)vs.(501.09±73.91),P<0.05]、CB2RmRNA[(4.00±0.19)vs.(2.99±0.28),P<0.05]、CB2R蛋白[(0.61±0.05)vs.(0.44±0.08),P<0.05]以及NLRP3[(4.75±0.40)vs.(2.53±0.26),P<0.05]、caspase-1[(8.76±0.72)vs.(6.02±0.35),P<0.05]、caspase-11[(16.31±1.13)vs.(11.43±0.83),P<0.05]、GSDMD[(16.46±1.22)vs.(10.46±0.62),P<0.05]mRNA表达升高。Objective To explore the correlation between cannabinoid 2 receptor(CB2R)and pyroptosis-related indicators in mice with septic lung injury.Methods Mice were randomly(ramdon number)divided into four groups(n=6 per group):sham operation group(sham),mild sepsis group(ALIMi),moderate sepsis group(ALIMo)and severe sepsis group(ALIS).The model of septic lung injury was established by cecal ligation and puncture.The wet-dry weight ratio of lung tissues and lung injury scores were measured 12 hours after operation.The expression of CB2R protein was measured by western blot,and the expression of mRNA of CB2R,NLRP3,caspase-1/11,GSDMD were detected by RT-PCR.Meanwhile ELISA was used to measure the level of inflammatory factor IL-6 and TNF-α.SPSS 22.0 software was used for data analysis.Multiple comparison was analyzed by one-way analysis of variance(one-way ANOVA)and comparison between two groups was performed by LSD test or Games-Howell test.Then,the correlation between the expression of CB2R mRNA and the level of inflammatory cytokines as well as the expression of the pyroptosis-related indicators mRNA was analyzed by pearson correlation analysis,respectively.Results The statistical value F was obtained by one-way ANOVA and comparison between two groups was performed.Compared to sham group,all above indicators increased with the aggravation of inflammation in the sepsis groups(P<0.05).Compared to ALIMi group,the concentrations of IL-6[(277.31±41.07)vs.(140.09±27.56),P<0.05]and TNF-α[(501.09±73.91)vs.(261.36±40.73),P<0.05]in lung tissue homogenate increased in ALIMo group.And the level of CB2mRNA[(2.99±0.28)vs.(2.02±0.19),P<0.05],the expression of CB2 protein[(0.44±0.08)vs.(0.23±0.05),P<0.05]and the level of NLRP3[(2.53±0.26)vs.(1.61±0.15),P<0.05],caspase-1[(6.02±0.35)vs.(3.60±0.38),P<0.05],caspase-11[(11.43±0.83)vs.(6.30±0.65),P<0.05]and GSDMD[(10.46±0.62)vs.(5.67±0.54),P<0.05]mRNA also increased.Compared to ALIMo group,the concentrations of IL-6[(475.90±67.65)vs.(277.31±41.07),P<0.05]and TNF-α[(713.9

关 键 词：大麻素2型受体 细胞焦亡 脓毒症 急性肺损伤 炎症 盲肠结扎穿孔法 mRNA 相关性 

分 类 号：R459.7[医药卫生—急诊医学] R563[医药卫生—治疗学]