Practical Journal of Organ Transplantation(Electronic Version) ›› 2016, Vol. 4 ›› Issue (1): 46-51.
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秦科 1 ,邝晓聪 2 ,孙煦勇 1 ,董建辉 1 ,蓝柳根 1 ,黄莹 1 ,曹嵩 1 ,李美思 1 ,李壮江 1,李海滨 1 ,苏庆东 1 ,周洁惠 1
Abstract:
Objective To observe the alterations of macrophage M1 and M2 subsets and the regulation of Ulinastatin(UTI)and to explore the mechanisms of UTI anti-inflammatory in the immunocompromised mousewith severe pulmonary infection. Methods A total number of 60 Balb/c mice were randomly divided into control group,model group and UTI-group,with 20 mice in each group. The mice in model group were prepared withmethylprednisolone by intraperitoneal injectionand dripping LPS bytrachea. The mice inUTI-group were treated by intervention with UTI(1×105U/kg),before or after LPSIntervention. At6 hours after theintratracheal administration of LPS,the indexes were detected as fellow: the cell types and ratios of cells in bron choalveolar lavage fluid(BALF),pathological changes expression of CD163 in the lung of mice were detected by hematoxylin-eosin(HE)staining and immunohistochemistry staining,respectively. The mRNA expression of M1-type markers(TNF-α,IL-6,iNOS)andM2-type markers(CD206,Arg-1,IL-10)were analyzed by real-time quantitative PCR in BALF. Results There was acute respiratory distress phenomenon in the model group. The pathological changes were wider alveolar walls and predominant polymorphonuclear neutrophils(PMN)and mononuclear inflammatory infiltrates. In UTI group,the pulmonary injury were alleviated and lung injury score were significantly lower than that of model group〔lung injury score(score):6.77±0.85 vs. 8.21±0.92,P < 0.05〕. The ratios of PMN and macrophage were significantlyincreased in in UTI group and model group,but there were no significant difference between the two groups. Except iNOS,the markers of M1 and M2 macrophages were higher than those of control group. In UTI group,M1-type markers(TNF-α,IL-6)and M2-type markers(CD206,IL-10)were significantly lower than that in model group〔TNF(2-ΔΔCt):0.42±0.18vs. 0.76±0.17,IL-6(2-ΔΔCt):0.72±0.26 vs. 1.22±0.32,IL-10(2-ΔΔCt):3.91±1.35vs. 6.84±2.13,CD206(2-ΔΔCt):0.61±0.19 vs.0.92±0.31,allP < 0.05〕,ad iNOSndArg-1hadnosignificant difference. In additional,M2-type marker CD163 expression in UTI group was significantly lower than that of model group,but higher than that of control group〔CD163(A)value :31 430.0±2 417.97 vs. 69 855.0±6 489.5,P < 0.05〕. Conclusion In the immune suppression mice of Th1 to Th2 of unidirectional deviation,M1 macrophage subset was activated more strongly than M2 macrophage subsets in severe pulmonary infection.The mechanisms of UTIanti-inflammatory may regulate of rebalance of M1 and M2 macrophages.
摘要:
目的 探讨免疫抑制状态下重症肺部感染小鼠肺巨噬细胞亚型变化情况,观察乌司他丁(UTI) 干预的巨噬细胞调控效应,初步明确其拮抗重症肺部炎症的作用机制。方法 选取 60 只 Balb/c 小鼠,分为对照组、模型组和乌司他丁组,每组 20 只小鼠 ;其中模型组采用腹腔注射甲泼尼龙(30 mg/kg),气管内滴注内毒素(10 mg/kg)制备免疫低下急性重症肺部感染小鼠模型 ;乌司他丁组则在建模前后腹腔注射乌司他丁(1×105 U/kg)干预 ;主要观察小鼠呼吸情况、肺泡灌洗液炎性细胞构成、肺组织病理改变与评分以及 M1 和 M2 巨噬细胞等指标,采用免疫组化法检测肺组织 M2 巨噬细胞的 CD163 表达,采用实时定量 PCR检测肺泡灌洗液细胞中 M1 巨噬细胞分泌分子肿瘤坏死因子 -α(TNF-α)、白细胞介素 -6(IL-6)、诱导型一氧化氮合成酶(iNOS)和 M2 巨噬细胞标志与分泌分子 CD206、Arg-1、IL-10 的 mRNA 表达水平。结果 模型组和乌司他丁组小鼠一般情况较差,部分小鼠出现呼吸窘迫现象,其中模型组小鼠肺泡腔与间质炎性细胞浸润,肺间质增宽,乌司他丁组病变减轻,与模型组相比,肺损伤评分显著降低〔(肺损伤评分(分):6.77±0.85 比 8.21±0.92,P < 0.05〕,乌司他丁组与模型组肺泡灌洗液中中性粒细胞比例显著增高,但两组差异不大。除 iNOS 外,M1 和 M2 巨噬细胞相关分子均高于对照组,与模型组相比,乌司他丁组 M1 巨噬细胞相关分子 TNF-α、IL-6 以及 M2 巨噬细胞相关分子 IL-10、CD206 均显著降低〔TNF(2-ΔΔCt):0.42±0.18 比 0.76±0.12,IL-6(2-ΔΔCt):0.72±0.26 比 1.22±0.32,IL-10(2-ΔΔCt):3.91±1.35 比 6.84±2.13,CD206(2-ΔΔCt):0.61±0.91 比 0.92±0.31,均 P < 0.05〕,而 iNOS 和 Arg-1 则与模型组无显著差别。此外,免疫组化染色检测乌司他丁组肺组织 M2 巨噬细胞的 CD163 表达,其结果明显低于模型组〔CD163(A)值 :31 430.0±2 417.97 比 69 855.0±6 489.5,P < 0.05〕,但高于对照组。结论 在辅助 T 细胞 Th1 单向偏移 Th2 免疫抑制状态中,重症肺部感染的巨噬细胞呈 M1 偏向 M2 亚群优势活化的状态,乌司他丁则可能通过调控 M1 与 M2 亚群再平衡来发挥其抗炎效应。
秦科, 邝晓聪, 孙煦勇, 董建辉, 蓝柳根, 黄莹, 曹嵩, 李美思, 李壮江, 李海滨, 苏庆东, 周洁惠 .
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https://syqgyz.magtechjournal.com/EN/Y2016/V4/I1/46