脂多糖对小鼠肾脏炎症因子、过氧化物酶体增殖物活化受体γ及其辅调节因子表达的影响  

Effects of intraperitoneal injection of LPS on expression of MCP-1,iNOS,PPAR-γ and related coregulators in mice kidneys

作　　者：陈慧[1] 韩琳[1] 仲芳[1] 王伟铭[1] 陈楠[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院肾脏科,上海200025

出　　处：《肾脏病与透析肾移植杂志》2008年第5期432-438,共7页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：国家自然科学基金资助项目(30270613;30771000);上海市重点学科(T0201);上海市卫生局重点学科基金(05III001);上海市卫生局重点课题(2003ZD002)

摘　　要：目的:观察腹腔注射脂多糖(LPS)对小鼠肾脏单核细胞趋化蛋白1(MCP-1)、诱导型一氧化氮合酶(iNOS)、过氧化物酶体增殖物活化受体γ(PPAR-γ)及其辅调节因子表达的影响。方法:雄性云南昆明小白鼠[(20±2)g]随机分成两组:LPS组,腹腔注射LPS(5mg/kg);对照组,腹腔注射等体积磷酸盐缓冲液。分别于0～72h后处死小鼠,检测血清尿素氮和肌酐变化;留取肾脏,EMSA法检测NF-κB及PPAR-γ的DNA结合活性;Real-time PCR法检测肾组织MCP-1、iNOS、PPAR-γ及其辅激活因子1(PGC-1)、类固醇受体辅激活因子1(SRC-1)、SRC-2、SRC-3及核辅抑制因子(NCoR)mRNA的表达;ELISA检测肾组织MCP-1蛋白表达,Western Blot检测肾组织核蛋白PPAR-γ表达;HE染色观察病理改变,免疫组织化学法观察肾组织巨噬细胞的浸润情况。结果:小鼠腹腔注射LPS后血清尿素氮升高明显(P<0.05),但血肌酐无明显变化。肾组织NF-κB的DNA结合活性在0.5h后即明显增高,而PPAR-γ的DNA结合活性早期增强,8h后开始减弱。与同时间点对照组及基础值相比,小鼠腹腔注射LPS后6h,12h及24h,肾组织MCP-1 mRNA及蛋白表达均显著增加(P<0.01);而iNOS mRNA表达在6h及12h后显著增加(P<0.01)。PPAR-γ及PGC-1 mRNA表达则显著下调(P<0.01),核内PPAR-γ蛋白含量早期升高,24h时显著下降(P<0.01)。SRC-1、SRC-2、SRC-3及NCoR mRNA的表达与对照组相比无显著差异。LPS作用后肾组织HE染色未见显著改变,免疫组化可见肾组织巨噬细胞浸润,最初主要分布在髓质肾小管周围,在48h及72h浸润更为明显,皮质肾小球周围也可见巨噬细胞浸润。结论:小鼠腹腔注射LPS后肾组织中NF-κB活性及相关炎症因子MCP-1和iNOS表达上调,肾组织内有明显的巨噬细胞浸润,表明处于炎症状态,而PPAR-γ及其辅激活因子PGC-1的表达下调可能与炎症发展相关。Objective：To observe the effects of hpopolysaccharide （LPS） by intraperitoneal injection on expression of MCP-1, iNOS, PPAR-γ and related coregulators in mice kidneys. Methodology：Male KM mice （weight：20±2g） were randomized to two groups. They were LPS group injected with LPS（5mg/kg） by intraperitoneal route and control group injected with equal volume of PBS. The mice were sacrificed after 0-72 hours, then blood urea nitrogen （BUN） and serum creatinine （SCr） levels were measured. The DNA binding activity of NF-KB and PPAR-γ, in kidneys was measured with electrophoretic mobility shift assay （EMSA）, and the mRNA expression of MCP-1, iNOS, PPAR-γ, PGC-1, SRC-1, SRC-2, SRC-3 and NCoR in kidneys were detected by real-time PCR. ELISA and Western blot were applied to analyze the protein expression of MCP-1 and PPAR-γ, respectively. The pathologic change and macrophage recruitment to the kidney were detected by HE and immunohistochemistry （IHC） staining. Results：The level of BUN was increased significantly after LPS injection（P 〈 0. 05 ）while SCr was no significant change between the two groups. The DNA binding activity of NF-KB was significantly enhanced at 0. 5h after intraperitoneal injection of LIPS, while the DNA binding activity of PPAR-γ was elevated at early time, then gradually down-regulated after 8 hours. Compared with control group and baseline, the mRNA and protein expression of MCP-1 in kidneys of mice was significantly up-regulated after 6, 12, and 24 hours （P 〈 0. 01 ）. The mRNA expression of iNOS was significantly up-regulated after 6 and 12 hours （ P 〈 0.01 ）. The mRNA expression of PPAR-γ and PGC-1 was down-regulated after 6 and 12 hours （P 〈 0.01 ）and the nuclear protein level of PPAR-3, was up-regulated at early time bat down-regulated at 24 hours （P 〈 0. 01 ）. The mRNA expression of SRC-1, SRC-2, SRC- 3 and NCoR was no significant changes. The morphological changes in the mice kidneys intraperitoneal injected with LPS were not

关 键 词：脂多糖 过氧化物酶体增殖物活化受体1 辅调节因子 炎症 

分 类 号：R692[医药卫生—泌尿科学]