慢性华支睾吸虫感染极化M2型巨噬细胞对脓毒症大鼠肺脏保护作用的研究  

Protective effects of chronic clonorchis sinensis infestation on lungs of septic rat via M2 macrophages polarization

作　　者：廖瑾莉[1] 熊艳[1] 刘志豪[1] 张婉婉[1] 祝锋[1] 詹红[1] 徐嘉[1] 

机构地区：[1]中山大学附属第一医院急诊科,广州510080

出　　处：《中华急诊医学杂志》2017年第5期533-537,共5页Chinese Journal of Emergency Medicine

基　　金：广东省医学科学技术研究基金（A2015486）;广东省科技计划项目（20128031800366）;广东省自然科学基金（$2013010015328）

摘　　要：目的初步探讨慢性华支睾吸虫（clonorchis sinensis，Cs）感染对脓毒症大鼠肺脏保护作用及其机制。方法经口Cs虫卵灌胃建立慢性Cs感染SD大鼠模型，细胞流式技术检测慢性Cs感染组和正常对照组腹腔巨噬细胞（Mφ）亚型，RT-PCR法检测Ⅰ型精氨酸酶（Arg-1）、发现炎症区域1（FIZZ 1）、诱导型一氧化氮合酶（iNOs）、肿瘤坏死因子（TNF-α）和白细胞介素-10（IL-10）mRNA的表达；结肠结扎穿孔术（cecal ligation and puncture，CLP）建立SD大鼠脓毒症模型，分为空白对照组、假手术组、脓毒症组，正常腹腔Mφ过继转移组、慢性Cs感染腹腔Mφ过继转移组和慢性Cs感染组。统计各组大鼠72 h累积病死率，观察肺脏病理变化，ELISA法检测CLP术后0、24、48和72 h血清TNF-α和IL-10的水平。结果正常大鼠腹腔Mφ以M1为主（91.9%），慢性Cs感染组大鼠腹腔Mφ则向M2极化（95.1%）。iNOS mRNA在M1中高表达，在M2中基本不表达；Arg-1、FIZZ 1和IL-10 mRNA在M2表达明显高于M1，TNF-α mRNA在M1中的表达高于M2（P〈0.05）。慢性Cs感染和M2腹腔过继转移组脓毒症大鼠的72 h病死率下降（P〈0.05），肺组织炎症损伤减轻，血清促炎因子TNF-α水平下降，抑炎因子IL-10水平上升（P〈0.05）。结论慢性Cs感染大鼠的M2型巨噬细胞极化可降低脓毒症时血清的促炎因子水平，提高抑炎因子水平，改善肺脏病理损伤，对脓毒症发挥保护作用。Objective To preliminarily investigate the protective effect of chronic clonorchis sinesis （Cs） infestation against sepsis in Sprague Dawley （SD） rats in order to explore its underlying mechanism. Methods Chronic Cs infestation model of SD rats was reproduced by intra-gastric administration with Cs ova. Twenty rats were randomly （random number） divided into normal group （n = 10） and Cs group （n = 10）. The proportion of differentiation in M1 and M2 macrophages were detected by flow cytometry. The expressions of Arg-1 （arginine-1）, FIZZ 1, iNOs and TNF-α mRNA were examined by reverse transcriptase polymerase chain reaction （RT-PCR） . The cecal ligation and puncture （CLP） procedure was performed to reproduce sepsis model of SD rats. Sixty rats were randomly （ random number） divided into control group, SHAM group, CLP group, Mφ ＋ CLP group, Cs-Mφ ＋ CLP group, and Cs-CLP group. The cumulative mortalities were calculated. The pathological changes of the lung tissue in different groups were demonstrated by HE staining. The serum levels of cytokines TNF-α and IL-10 were detected by ELISA at 0, 24, 48 and 72 h after CLP procedure. Results Compared with M1 peritoneal macrophages differentiation in control group （91.9%）, rat peritoneal macrophages were activated to M2 differentiation （95. 1% ） in chronic Cs infection group. RT-PCR assay showed expression of Arg-1 and FIZZ 1 mRNA were higher in M2 macrophages, and on the contrary, the expression of iNOS mRNA expression was higher in M1 macrophages. The expression of TNF-α mRNA in M1 was significantly higher than that in M2, whereas the expression of IL-10 mRNA in M2 was higher than that in M1. The cumulative mortality of septic rats 72 h after CLP procedure were much lower in both chronic Cs infestation group and M2 macrophages adoptive transfer group （ CLP group 70% vs. Mφ＋ CLP group 50% vs. Cs-Mφ ＋ CLP group 30% vs. Cs-CLP group 0%, P 〈 0.05 ） . In these two groups, the pathological damages in lung tissues we

关 键 词：脓毒症 慢性华支睾吸虫感染 替代活化型(M2)巨噬细胞 细胞因子 基因表达 

分 类 号：R459.7[医药卫生—急诊医学]