NecroX-5调节M1巨噬细胞极化减轻急性呼吸窘迫综合征肺损伤的机制  

Mechanisms of NecroX-5 regulating the polarization of M1 macrophages to reduces lung injury in acute respiratory distress syndrome

作　　者：朱丽华[1] 章忠明[2] 李薇[1] 冯霞 王芃[1] 刘兰燕 ZHU Lihua;ZHANG Zhongming;LI Wei;FENG Xia;WANG Peng;LIU Lanyan(Department of Physical Examination and Health Management,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院体检部健康管理部,广西南宁530021 [2]广西医科大学第一附属医院血液内科,广西南宁530021 [3]中山大学附属东华医院传染病科,广东东莞523110 [4]广西医科大学基础医学院,广西南宁530021

出　　处：《皖南医学院学报》2021年第3期205-209,共5页Journal of Wannan Medical College

基　　金：广西医疗卫生适宜技术开发与推广应用项目(S2018009)。

摘　　要：目的:探讨NecroX-5对脂多糖(LPS)诱导的急性呼吸窘迫综合征(ARDS)大鼠肺损伤的影响及机制。方法:将40只SD大鼠随机分为对照组、NecroX-5组、LPS组和NecroX-5+LPS组。采用尾静脉注射LPS诱导ARDS模型,NecroX-5组和NecroX-5+LPS组造模前7 d腹腔注射10 mg/kg NecroX-5。造模后24 h收集大鼠肺泡灌洗液(BALF),ELISA检测TNF-α、IL-6、IL-23、iNOS水平,HE染色检测肺组织病理变化,免疫组化染色检测肺组织CD68^(+)CD86^(+)表达,qPCR检测CD86、iNOS mRNA表达,Western blot法检测CD86、iNOS、HMGB1、RAGE蛋白表达。RAW264.7细胞随机分为对照组、NecroX-5组、LPS+IFNγ组和NecroX-5+LPS+IFNγ组,流式细胞术检测各组巨噬细胞极化情况,Western blot法检测HMGB1、RAGE蛋白表达。结果:与LPS组比较,NecroX-5+LPS组BALF中TNF-α、IL-6、IL-23、iNOS含量下降,肺组织损伤程度减小,CD68^(+)CD86^(+)阳性表达降低,iNOS、CD86 mRNA与蛋白表达量下降,HMGB1、RAGE蛋白表达下降(P<0.05)。与LPS+IFNγ组比较,NecroX-5+LPS+IFNγ组细胞F4/80+CD11b+CD11c+标记百分比下降,此外,HMGB1与RAGE蛋白表达下降(P<0.05)。结论:NecroX-5可能通过调节M1型巨噬细胞极化与HMGB1/RAGE表达,改善大鼠ARDS肺损伤。Objective:To investigate the effect and mechanisms of NecroX-5 on lung injury in rat models from acute respiratory distress syndrome(ARDS)induced by LPS.Methods:Forty SD rats were randomly divided into control group,NecroX-5 group,LPS group and NecroX-5+LPS group.LPS was used to induce ARDS model via tail vein injection.Rats in NecroX-5 and NecroX-5+LPS groups were intraperitoneally injected with NecroX-5 in dose of 10 mg/kg at the 7th day before modeling,and the rat alveolar lavage fluid(BALF)was collected at 24 h after modeling.The levels of TNF-α,IL-6,IL-23 and iNOS were detected by ELISA,and HE staining was performed to examine the pathological changes in the lung tissues.Immunohistochemistry was used to detect the expression of CD68^(+)CD86^(+)in the lung tissues.mRNA expression in CD86 and iNOS was detected with qPCR,and expression of CD86,iNOS,HMGB1 and RAGE was measured with Western blot.Then RAW264.7 cells were randomly divided into control group,NecroX-5 group,LPS+IFNγgroup and NecroX-5+LPS+IFNγgroup,and observed for the polarization of macrophages in each group by flow cytometry.Western blot was performed to detect the protein expression of HMGB1 and RAGE.Results:Compared with LPS group,levels of TNF-α,IL-6,IL-23 and iNOS in the BALF of the NecroX-5+LPS group as well as the degree of lung tissue damage were decreased and reduced,and the positive expression of CD68^(+)CD86^(+),the mRNA and protein expression of iNOS and CD86 decreased as well as the protein expression of HMGB1 and RAGE were all decreased(P<0.05).The percentage of F4/80+CD11b+CD11c+markers,together with the protein expression of HMGB1 and RAGE was decreased in the NecroX-5+LPS+IFNγgroup compared to LPS+IFNγgroup(P<0.05).Conclusion:NecroX-5 may improve the lung injury of rats from acute respiratory distress syndrome by regulating the polarization of M1 macrophages and the expression of HMGB1/RAGE.

关 键 词：急性呼吸窘迫综合征 NecroX-5 M1巨噬细胞极化 HMGB1/RAGE途径 肺损伤 

分 类 号：R563.8[医药卫生—呼吸系统]