丝裂原活化蛋白激酶激活的蛋白激酶2基因在白色念珠菌诱导急性肺损伤中的作用及其机制  

Role and mechanism of mitogen-activated protein kinase-activated protein kinase 2 gene in acute lung injury induced by Candida albicans

作　　者：袁润林[1] 俞谦[1] 杨仁保[2] 陈正徐[1] Yuan Runlin;Yu Qian;Yang Renbao;Chen Zhengxu(Clinical Laboratory of The Second People's Hospital of Hefei,Hefei(Hospital Affiliated to Anhui Medical University),Hefei 23001l,China;Department of General Surgery,Second People's Hospital of Hefei,Hefei(Hospital Affiliated to Anhui Medical University),Hefei 230011,China)

机构地区：[1]合肥市第二人民医院(安徽医科大学附属合肥医院)检验科,合肥230011 [2]合肥市第二人民医院(安徽医科大学附属合肥医院)普外科,合肥230011

出　　处：《中华实验外科杂志》2024年第10期2260-2264,共5页Chinese Journal of Experimental Surgery

基　　金：2020年度安徽高校自然科学研究重点项目(KJ2020A02170)。

摘　　要：目的:丝裂原活化蛋白激酶激活的蛋白激酶2(MK2)基因在白色念珠菌(CA)诱导急性肺损伤中的作用及机制。方法:野生型C57/b6小鼠分为MK2组和MK2+CA组;MK2基因敲除C57/b6小鼠分为MK2 KO组和MK2 KO+CA组;MK2+CA组和MK2 KO+CA组小鼠麻醉后经气管注射50μl白色念珠菌,MK2组和MK2 KO组小鼠麻醉后器官注射50μl生理盐水。分析4组小鼠存活情况;采用苏木精-伊红染色分析炎症情况;采用酶联免疫吸附试验(ELISA)和荧光定量聚合酶链反应(qPCR)分析血清炎性因子白细胞介素(IL)-6、IL-1β、肿瘤坏死因子(TNF)-α蛋白和mRNA水平;分离肺组织,采用蛋白质免疫印迹分析肺组织磷酸化p55和p65表达水平;采用流式细胞术分析巨噬细胞激活情况。组间比较采用t检验。结果:MK2+CA组肺病理损伤评分(8.74±1.54)明显高于MK2 KO+CA组小鼠(3.98±0.99),差异有统计学意义(t=5.412,P<0.05)。MK2组和MK2 KO组小鼠肺组织湿重/干重比值(3.39±0.15、3.42±0.20)比较,差异无统计学意义(t=0.267,P>0.05)。MK2 KO组小鼠肺组织湿重/干重比值(3.42±0.20)明显低于MK2 KO+CA组(4.32±0.20),差异有统计学意义(t=9.439,P<0.05)。MK2+CA组小鼠肺组织湿重/干重比值(5.79±0.21)明显高于MK2 KO+CA组(4.32±0.20),差异有统计学意义(t=14.300,P<0.05)。MK2+CA组小鼠血清炎性因子TNF-α、IL-6、IL-1β水平[(76.84±7.57)、(97.47±8.85)、(51.63±4.24)ng/ml]明显高于MK2 KO+CA组[(51.90±6.51)、(42.67±4.65)、(28.63±3.35)ng/ml],差异有统计学意义(t=7.058、15.500、12.030,P<0.05)。MK2+CA组小鼠血清F4/80+/CD11b+/CD80+巨噬细胞比例[(87.26±4.70)%]明显高于MK2 KO+CA组[(53.05±10.95)%],差异有统计学意义(t=8.120,P<0.05)。MK2+CA组小鼠肺组织肺组织磷酸化p65和p50蛋白表达水平(1.82±0.10、1.33±0.06)明显高于MK2 KO+CA组(1.23±0.10、1.00±0.07),差异有统计学意义(t=11.780、9.810,P<0.05)。结论:MK2参与白色念珠菌诱导急性肺损伤进程,敲除MK2可显著缓解巨噬细胞激Objective:Mitogen-activated protein kinase-activated protein kinase 2(MK2)gene has been detected in Candida albicans.Study on the role and mechanism of CA in inducing acute lung injury.Methods:Wild type C57/b6 mice were divided into MK2 group and MK2+CA group.MK2 knockout C57/b6 mice were divided into MK2 KO group and MK2 KO+CA group.Mice in MK2+CA group and MK2 KO+CA group were injected with 50μl Candida albicans through trachea after anesthesia,and mice in MK2 group and MK2 KO group were injected with 50μl normal saline after anesthesia.The survival of four groups of mice was analyzed.Hematoxylin-eosin staining was used to analyze the inflammation.Enzyme-linked immunosorbent assay(ELISA)and real time fluorescence quantitative polymerase chain reaction(PCR)were used.The levels of serum inflammatory factors interleukin-6,interleukin-1β,tumor necrosis factor a protein and mRNA were analyzed by qPCR.Lung infiltrating macrophages were isolated and the expression levels of phosphorylated p55 and p65 were analyzed by western blot.Macrophage activation was analyzed by flow cytometry.The measurement between groups was compared by t test.Results:The lung pathological injury score in the MK2+CA group(8.74±1.54)was significantly higher than that in the MK2 KO+CA group mice(3.98±0.99,t=5.412,P<0.05).The wet weight/dry weight ratio of MK2 group and MK2 KO group was 3.39±0.15,3.42±0.20,the difference was not statistically significant(t=0.267,P>0.05).The wet weight/dry weight ratio of MK2 KO group(3.42±0.20)was significantly lower than that of MK2 KO+CA group(4.32±0.20),and the difference was statistically significant(t=9.439,P<0.05).The lung wet weight/dry weight ratio in MK2+CA group(5.79±0.21)was significantly higher than that in MK2 KO+CA group(4.32±0.20),and the difference was statistically significant(t=14.300,P<0.05).Levels of serum inflammatory factors tumor necrosis factor a,interleukin-6 and interleukin-1βin MK2+CA group[(76.84±7.57),(97.47±8.85),(51.63±4.24)ng/ml]was significantly higher MK2 KO+CA gro

关 键 词：丝裂原活化蛋白激酶激活的蛋白激酶2 白色念珠菌 急性肺损伤 巨噬细胞 

分 类 号：R519.3[医药卫生—内科学]