IL-10对于耐药BCG感染巨噬细胞后iNOS和乳酸表达水平的影响  

Effects of IL-10 on the expression of iNOS and lactic acid in macrophages infected by drug-resistance BCG

作　　者：庞慧 戎燕筱[3] 马翠卿 张洵[2] 王家超 高雪[1] 魏林[1] PANG Hui;RONG Yanxiao;MA Cuiqing;ZHANG Xun;WANG Jiachao;GAO Xue;WEI Lin(Department of Immunology,Hebei Medical University,Shijiazhuang 050017,China;Department of Medical Laboratory Science,Changzhi Medical College,Changzhi 046000,China;Department of Infection,Shijiazhuang Fifth Hospital,Shijiazhuang 050017,China)

机构地区：[1]河北医科大学免疫学教研室,石家庄050017 [2]长治医学院医学检验学系,046000 [3]石家庄市第五医院感染科,050017

出　　处：《免疫学杂志》2023年第5期396-402,共7页Immunological Journal

基　　金：山西省基础研究计划(自由探索类)(20210302124326);山西省卫生健康委科研课题(2021005);河北省重点研发计划项目(20377716D);山西省高校科技创新项目(201802092);长治医学院学术技术带头人项目(XSQ201901);国家级大学生创新创业训练计划项目(20220870);山西省大学生创新创业训练计划项目(20220870)。

摘　　要：目的通过IL-10作用于耐药BCG感染巨噬细胞的体外模型,研究其对巨噬细胞产生iNOS和乳酸的影响,为筛选耐药分枝杆菌诊疗靶点提供理论依据。方法采用Western blot检测巨噬细胞中STAT1、STAT3表达水平;运用比色法测定诱导型一氧化氮合酶(iNOS)及乳酸含量;通过小干扰RNA敲低IL-10,分析STAT1、STAT3、iNOS、乳酸等表达水平。结果敲除IL-10与未敲除IL-10的2组细胞在敏感BCG(S’与S组)作用下,S’组诱导STAT1的能力受到影响;耐药BCG(R’与R组)诱导STAT1的能力较敏感BCG更弱;而STAT3在R’组显著低于S’组(P=0.0018);R’组比R组也有显著降低(P=0.0041)。敲低IL-10后,iNOS、乳酸在R’组表达水平均显著升高。结论耐药BCG促进巨噬细胞向M2型分化以及产生杀菌酶的能力依赖于IL-10。This study was designed to evaluate the effect of IL-10 on drug-resistance BCG acting on the production of inducible nitric oxide synthase(iNOS)and lactic acid by macrophages in vitro,providing the theoretical basis on screening the diagnosis and treatment of drug-resistance mycobacteria.The expression of STAT1 and STAT3 in macrophages were detected by Western blotting;the content of iNOS and lactic acid was determined by colorimetry.The levels of STAT1,STAT3,iNOS and lactic acid were also analyzed after knocking down IL-10 with small interfering RNA(si-IL-10).Data showed that under the condition of drug-sensitive BCG(S'and S groups),STAT1 was influenced in the group S'with si-IL-10 or not,while drug-resistance BCG had weaker ability to induce STAT1 contrast with drug-susceptibility one;and STAT3 in the R'group was remarkably less than that in the S'group or R group(P=0.0018;P=0.0041).After si-IL-10,both of iNOS and lactic acid were ascended in the R'group.In conclusion,the capacity of drug-resistance BCG to promote macrophage to differentiate into M2 and produce bactericidal enzymes are depended on IL-10.

关 键 词：耐药BCG 白介素-10 巨噬细胞 诱导型一氧化氮合酶 乳酸 

分 类 号：R392.1[医药卫生—免疫学]