NF-κB及下游炎症因子在胰岛素抵抗大鼠肾脏中的表达  

Expressions of NF-κB and downstream inflammatory factors in the kidney of insulin resistance rat

作　　者：闫双通[1] 李春霖[1] 陆菊明[2] 田慧[1] 谷昭艳[1] 刘瑜[1] 

机构地区：[1]解放军总医院老年内分泌科,北京100853 [2]解放军总医院内分泌,北京100853

出　　处：《解放军医学杂志》2014年第10期782-786,共5页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81173625,30873412,30801201)~~

摘　　要：目的探讨胰岛素抵抗(IR)大鼠肾脏组织中核转录因子(NF-κB)及下游炎症因子诱导型一氧化氮合酶(iNOS)、环氧合酶(COX-2)表达的变化及意义。方法健康雄性Wistar大鼠30只,随机分为正常对照(NC)组(n=15)、IR组(n=15)。以高脂高糖喂养法构造IR大鼠模型,并使用高胰岛素正葡萄糖钳夹实验验证模型的有效性。留取大鼠肾脏组织,HE染色观察肾脏结构的变化。免疫组化染色观察肾脏组织中iNOS和COX-2的表达情况。RT-PCR检测肾脏组织中NF-κB、iNOS和COX-2 mRNA的表达情况。采用凝胶电泳迁移率(EMSA)法测定大鼠肾脏组织NF-κB的DNA结合活性。结果 HE染色结果显示,与NC组相比,IR组大鼠可见肾小球体积增大、系膜区增宽等肾脏组织早期受损表现。免疫组化染色结果显示,与NC组比较,IR组大鼠肾脏组织中iNOS和COX-2表达水平明显升高(P<0.05)。RT-PCR结果显示,IR组大鼠肾脏组织中NF-κB mRNA及iNOS mRNA、COX-2 mRNA表达水平与NC组比较均明显升高(P<0.05)。EMSA结果显示,IR组大鼠肾脏组织核蛋白NF-κB DNA结合力与NC组比较明显升高(P<0.05)。结论 IR大鼠肾脏组织中存在NF-κB的激活,从而启动下游靶基因iNOS、COX-2表达的增加,进而导致肾脏组织受损。NF-κB的激活可能是导致IR大鼠肾脏病变的始动因子之一。Objective To investigate the variation and significance of the expressions of NF-κB, inducible nitric oxide synthase（iNOS） and cyclooxygenase-2（COX-2） in the renal tissue of insulin-resistant rat. Methods Thirty healthy male Wistar rats were bred since 2 months old, and they were randomly divided into normal control（NC） group（n=15） and insulin-resistant（IR） group（n=15）. Insulin resistance rat model was reproduced by feeding with high fat and sucrose diet. Hyperinsulinemic-euglycemic clamp test was used to verify the reproduction of the model. The kidneys of the rats were obtained after the successful reproduction of the model. The change in renal histology was observed by HE staining, and the expressions of iNOS and COX-2 in the kidneys were detected by immunohistochemistry staining. The mRNA expressions of NF-κB, iNOS and COX-2 in the kidneys were assessed with RT-PCR. DNA binding activity of NF-κB in the rat＇s kidney was assessed with electrophoretic mobility shift assay（EMSA）. Results HE staining showed that, compared with NC group, the early lesions of the renal tissue, such as glomerular enlargement and mesangial region broadening, could be seen in IR group. Immunohistochemical staining showed that the positive expressions of iNOS and COX-2 were up-regulated significantly in IR group than in NC group（P〈0.05）. RT-PCR revealed that the expressions of NF-κB mRNA, iNOS mRNA and COX-2 mRNA in renal tissue were significantly higher in IR group than in NC group（P〈0.05）. EMSA showed that the binding activity of NF-κB in renal tissue increased significantly in IR group than in NC group（P〈0.05）. Conclusion NF-κB activation is present in the kidney tissue in the insulin resistance rat, which may upregulate the expression of downstream target gene iNOS and COX-2, resulting in damage to kidney tissue. The activation of NF-κB may be one of the initiative factors that lead to the kidney lesion of the insulin resistance rat.

关 键 词：胰岛素抗药性 环氧化酶2 大鼠 肾 

分 类 号：R587.1[医药卫生—内分泌]