旋毛虫排泄分泌抗原53-ku重组蛋白上调M2型肺泡巨噬细胞减轻脓毒症小鼠急性肺损伤  

Recombinant Trichinella Spiralis Excretory-secretory Antigen 53ku Protein Alleviates Acute Lung Injury in Septic Mice via Polarization of M2 Macrophages

作　　者：关开泮[1] 江仁 徐雪 刘志豪[3] 廖瑾莉[3] 熊艳[3] 詹红[3] 曾庆理[3] 徐嘉[3] GUAN Kai-pan JIANG Ren XU Xue LIU Zhi-hao LIAO Jin-li XIONG Yan ZHAN Hong ZENG Qing-li XU Jia(Department of Intensive Care Unit, Division of Huiya Department of Internal Medicine, The Division of Huangpu Department of Emergency, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China)

机构地区：[1]中山大学附属第一医院惠亚院区重症监护室,广东广州510080 [2]中山大学附属第一医院黄埔院区内科,广东广州510080 [3]中山大学附属第一医院急诊科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2017年第5期670-675,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省医学科学技术研究基金(A2015486;B2014110);广东省自然科学基金(2016A030313249);广东省科技计划项目(2014A020212150)

摘　　要：【目的】探讨重组旋毛虫排泄分泌抗原53-ku蛋白(rTsP53)对小鼠急性肺损伤的保护作用及其机制。【方法】采用尾静脉脂多糖注射(LPS,10 mg/kg)建立BALB/c小鼠急性肺损伤模型。在肺损伤后2 h尾静脉注射rTsP53(50μg/只)治疗。统计各组小鼠72 h累计死亡率,HE染色法观察各组小鼠肺脏病理损伤情况,测量肺脏湿/干质量比(W/D),ELISA法检测各组小鼠肺泡灌洗液IL-6和IL-4浓度,RT-PCR法检测各组小鼠肺泡灌洗巨噬细胞TNF-α、iNOS、IL-10和Arg-1的mRNA表达。【结果】经rTsP53蛋白治疗可明显降低急性肺损伤小鼠72 h死亡率,减轻脓毒症小鼠肺脏组织的病理损伤,降低肺脏的湿干重比,降低Smith评分。LPS组小鼠肺泡灌洗液IL-6较空白组升高,而IL-4下降,灌洗巨噬细胞的TNF-α,iNOS的mRNA表达水平较空白组明显升高,而IL-10,Arg-1的mRNA表达水平下降,表现出M1表型极化的特点;经rTsP53蛋白治疗的脓毒症小鼠肺泡灌洗液IL-6浓度下降,IL-4水平上升,灌洗巨噬细胞向M2表型极化,表现为IL-10,Arg-1的mRNA表达水平较脓毒症组明显升高,而iNOS的mRNA表达水平下降。【结论】rTsP53蛋白通过上调M2型巨噬细胞抑制炎症反应和修复组织损伤,对脓毒症小鼠急性肺损伤发挥保护效应。【Objectives】To investigate the protective effects of recombinant Trichinella spiralis excretory-secretory 53 ku pro-tein（rTsP53）on acute lung injuries in mice.【Methods】Thirty Balb/c mice were randomly divided into normal group. ALI group andrTsP53 group（n = 10,respectively）. Macrophages were harvested by bronchoalveolar lavage. Mortality in 72 hours was counted andcompared. Pathological damage of lung tissues was observed by HE staining and graded by Smith score. Wet/dry ratio was measured.The bronchoalveolar lavage fluid concertration of IL-6 and IL-4 was measured by ELISA. The mRNA expression of TNF-α,iNOS,IL-10 and Arg-1 in alveolar lavage macrophages was detected by RT-PCR.【Results】72 h mortality of ALI mice was 70%,whichwas reduced to 30% in mice received rTsP53 treatment. Compared with ALI mice,the pathological damage of in rTsP53 treated-mice was improved and Smith score was declined,combined with descending W/D ratio. IL-6 level of alveolar lavage fluid waselevated in ALI mice compared with normal group. And alveolar lavage macrophage was polarized to M2 sub-type,appeared ashigher mRNA expression of TNF-α and iNOS and lower level of IL-10 and Arg-1. Bronchoalveolar lavage fluid concentration ofIL-6 was declined and IL-4 was elevated in rTsP53-treated mice compared with ALI group. The macrophages of alveolar wash hadhigher mRNA expression of IL-10 and Arg-1,while lower level of TNF-α and iNOS,manifesting M2 polarization characteristics.【Conclusion】Recombinant T.spiralis P53 protein could protect mice from acute lung injuries induced by LPS via modulating M2 macrophage polarization,which play a role in depression of inflammatory reaction and tissue repairment.

关 键 词：重组旋毛虫排泄分泌抗原53-ku蛋白 脂多糖 急性肺损伤 M2型巨噬细胞 

分 类 号：R392.1[医药卫生—免疫学]