尿毒症大鼠肠道巨噬细胞向促炎型分化加重微炎症状态研究  

Differentiation of intestinal macrophages towards proinflammatory phenotype aggravated the microinflammatory status in uremia rats

作　　者：孙凌霜[1] 刘华[1] 魏萌[1] 梁珊珊[1] 刘思秀 薛瑾虹 史珂慧[1] 蒋红利[1] SUN Ling-shuang;LIU Hua;WEI Meng;LIANG Shan-shan;LIU Sixiu;XUE Jin-hong;SHI Ke-hui;JIANG Hong-li(Department of Blood Purification, Nephrology Hospital,The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China)

机构地区：[1]西安交通大学第一附属医院肾脏病医院血液净化科

出　　处：《中国血液净化》2019年第6期406-410,共5页Chinese Journal of Blood Purification

基　　金：国家自然科学基金(81370838)

摘　　要：目的探讨尿毒症大鼠肠道巨噬细胞功能及其与尿毒症微炎症状态的关系。方法SD 大鼠随机分为:5/6 肾切除手术建立的尿毒症组和假手术组。检测血中内毒素、C 反应蛋白(C reactiveprotein,CRP)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平。透射电子显微镜观察肠巨噬细胞形态;检测肠组织切片中巨噬细胞活化指标CD11a 和诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)表达;realtime-PCR、Western Blot 法检测巨噬细胞表面的早期生长因子1(early growth response gene 1,EGR1)及Toll 样受体4(Toll-likereceptor 4,TLR4)mRNA 和蛋白的表达;检测肠组织切片中细胞间粘附分子1(intercellular adhesionmolecule-1,ICAM-1)、TGF-β的表达。结果尿毒症组与假手术组相比血中内毒素、IL-6、TNF-α和CRP明显增加水平显著增加(t 值分别为- 5.145,- 3.776,- 3.553,- 10.468;P 值分别为<0.001,0.001,0.002,<0.001)。尿毒症组的巨噬细胞突触和伪足较假手术组少。尿毒症大鼠肠组织内CD11a 和iNOS染色表达多;EGR1 及TLR4mRNA 和蛋白表达多;ICAM-1、转化生长因子-β(transforming growth factor,TGF-β)在尿毒症组表达多。结论尿毒症状态下肠道巨噬细胞向M1 型活化;巨噬细胞吞噬功能障碍;巨噬细胞炎症级联反应通过脂多糖激活EGR1 及TLR4,引起iNOS、TGF-β表达增多,血中CRP、IL- 6、TNF-α水平增加,加重微炎症状态。Objective To investigate the relationship between the function of intestinal macrophages and the microinflammation in uremic rats. Methods Male Sprague-Dawley rats were randomly divided into sham surgery group and uremia group. Blood levels of endotoxin, hs-CRP, IL-6 and TNF-α were assayed. The ultrastructure of macrophages was examined by transmission electron microscopy. Immunochemistry of the intestinal sections was used to analyze the expression of CD11a and iNOS, the active markers of macrophages, as well as the expression of intercellular adhesion molecule-1 (ICAM-1) and TGF-β. Quantitative real-time PCR and western blot were employed to assess the mRNA and protein levels of early growth response gene 1 (EGR1) and toll-like receptor 4 (TLR4). Results The blood levels of endotoxin, IL-6, TNF-α and hs-CRP were significantly higher in uremia group than in sham surgery group (t=-5.145,-3.776,-3.553 and -10.468 respectively;P=<0.001, 0.001, 0.002 and <0.001 respectively). The macrophages in uremia group showed fewer cytoplasmic protrusions and pseudopodia. The intestinal macrophages in uremia group exhibited heavier staining of CD11a, iNOS, ICAM-1 and TGF-β, and higher mRNA and protein levels of EGR1 and TLR4. Conclusions In uremia rats, intestinal macrophages differentiated towards proinflammatory phenotype, which was related to the microinflammatory status of the rats.

关 键 词：巨噬细胞 微炎症 尿毒症 

分 类 号：R692.5[医药卫生—泌尿科学]