印苦楝内酯靶向RNF114治疗金黄色葡萄球菌急性肺炎  

Nimbolide targets RNF114 in treatment of mouse model of acute pneumonia caused by Staphylococcus aureus

作　　者：孙慧 田湉 熊廷蓉 邹全明 张笑恺 杨宪 王于 SUN Hui;TIAN Tian;XIONG Tingrong;ZOU Quanming;ZHANG Xiaokai;YANG Xian;WANG Yu(Engineering Research Center of Active Matter Biotechnology of Ministry of Education,College of Life Sciences,Chongqing Normal University,Chongqing,401331;Department of Microbiology and Biochemical Pharmacy,National Engineering Research Center of Immunological Products,Faculty of Pharmacy and Laboratory Medicine,Army Medical University(Third Military Medical University),Chongqing,400038;Department of Basic Courses,Noncommissioned Officer School,Army Medical University(Third Military Medical University),Shijiazhuang,Hebei Province,050081,China)

机构地区：[1]重庆师范大学生命科学学院活性物质生物技术教育部工程研究中心,重庆401331 [2]陆军军医大学药学与检验医学系微生物与生化药学教研室,国家免疫生物制品工程技术研究中心,重庆400038 [3]陆军军医大学士官学校基础部,河北石家庄050081

出　　处：《陆军军医大学学报》2024年第12期1353-1360,共8页Journal of Army Medical University

基　　金：国家自然科学基金(82201942)。

摘　　要：目的探究印苦楝内酯(nimbolide,NIM)治疗金黄色葡萄球菌急性肺炎作用机制。方法通过气管插管构建小鼠金葡菌急性肺炎模型,NIM治疗后,观察小鼠生存率差异,平板培养分析肺部荷菌量变化,ELISA检测肺组织炎性细胞因子表达差异;ELISA及Western Blot检测腹腔原代巨噬细胞感染金葡菌后,NIM对炎症细胞因子的表达水平及炎症通路激活的影响;慢病毒shRNA敲低原代腹腔巨噬细胞RNF114蛋白水平,通过ELISA及Western Blot探究NIM是否通过RNF114抑制炎症反应。结果金葡菌感染小鼠肺部后可导致急性死亡,NIM显著提高了小鼠生存率,下调小鼠肺部炎症细胞因子水平,但短期内不影响金葡菌肺部定植;巨噬细胞感染结果表明,NIM可能通过调控RNF114功能,下调ERK的磷酸化水平抑制MAPK通路激活,进而抑制炎症细胞因子的表达。结论NIM可能通过调控巨噬细胞RNF114功能,抑制MAPK通路激活进而抑制小鼠肺部炎症细胞因子的表达,最终抑制金葡菌急性肺部高炎症导致的小鼠死亡。Objective To explore the mechanism which drives nimbolide(NIM)in treating acute pneumonia caused by Staphylococcus aureus(S.auteus).Methods A mouse model of acute pneumonia caused by S.auteus was constructed through endotracheal intubation.After NIM treatment,the survival rate was observed,the amount of bacteria in the lung was tested by plate culture,and the expression of inflammatory cytokines in the lung tissues was detected with ELISA.After primary cultured peritoneal macrophages(PM)were infected with S.auteus,the effect of NIM on the expression of inflammatory cytokines and activation of inflammatory pathway were studied with ELISA and Western blotting,respectively.The effect of RNF114 knockdown by lentiviral shRNA infection on inflammation responses in PM was explored with ELISA and Western blotting.Results Acute infection of S.auteus in the lung could cause acute death in the mice,while NIM treatment significantly improved the survival rate and down-regulated the levels of inflammatory cytokines in the lung.However,it had no effect on the lung colonization of S.auteus in the short term.The results of in vitro experiments indicated that NIM may regulate RNF114 function to down-regulate the phosphorylation level of ERK,inhibit the activation of MAPK pathway,and thus suppress the expression of inflammatory cytokines.Conclusion NIM may inhibit the activation of MAPK pathway by regulating the function of RNF114,and thus suppress the expression of inflammatory cytokines in the lung,and finally inhibit the death of mice with acute pulmonary hyperinflammation caused by S.auteus.

关 键 词：金黄色葡萄球菌 Nimbolide 巨噬细胞 RNF114 

分 类 号：R282.71[医药卫生—中药学] R378.11[医药卫生—中医学] R563.1