摘要
目的通过观察糖尿病(DM)大鼠肾组织中Notch2、含pyrin结构域NOD样受体家族3(NLRP3)的表达情况及α硫辛酰胺(ALM)对Notch2、Toll样受体4(TLR4)、NLRP3和胶原蛋白表达的影响,初步探讨ALM抗糖尿病肾组织炎症和肾纤维化的可能保护机制。方法以链脲佐菌素(STZ)复制DM大鼠模型,分为DM组和ALM组,同时设正常对照组(NC组),每组8只。DM造模成功2周后,ALM组用ALM[150 mg/(kg·d)]进行灌胃,治疗6周后处死大鼠。HE和Masson染色观察肾组织形态变化,免疫组织化学染色观察肾组织中Notch2、TLR4、NLRP3的表达和定位,Western blot法检测肾组织中Notch2、TLR4、NLRP3和4型胶原蛋白(Col4)的蛋白水平,ELISA检测肾组织匀浆中炎性因子白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)的水平。采用Spearman秩相关检验对Notch2与TLR4、NLRP3蛋白表达进行相关性分析。结果 DM组的血糖、24 h尿蛋白量、肾脏指数、总胆固醇、甘油三酯均显著高于NC组;与DM组相比,ALM组肾脏指数、24 h尿蛋白量明显降低,而血糖无明显改变。与NC组相比,DM组大鼠肾组织中Notch2、TLR4、NLRP3和Col4的蛋白水平增加,ALM组大鼠肾组织中的上述蛋白水平明显低于DM组; ALM组大鼠肾组织匀浆IL-6和TNF-α的水平较DM组明显降低。相关性分析显示在DM组以及ALM组中Notch2与TLR4的表达均呈显著正相关,Notch2与NLRP3的表达也均呈显著正相关。结论 ALM抑制DM大鼠肾组织的炎症和Col4的产生,与Notch2抑制TLR4和NLRP3的活化有关。
Objective To observe the expression of Notch2,NLR family pyrin domain containing 3( NLRP3) in the renal tissue of rats with diabetes mellitus( DM),and the effects of alpha-lipoamide( ALM) on the expression of Notch2,Toll-like receptor 4( TLR4),NLRP3 and collagen,to explore the possible protective mechanism of ALM against diabetic renal inflammation and renal fibrosis. Methods The DM rat models were induced by the injection of streptozotocin( STZ) and divided into DM group and ALM group. Meanwhile,normal control( NC) group was set up. There were 8 rats in each group.Two weeks after the successful establishment of DM model,ALM [150 mg/( kg·d) ]was given intragastrically to the rats in the ALM group,and the rats were sacrificed 6 weeks later. The morphologic changes of renal tissues were observed by HE and Masson staining. The expression and localization of Notch2,TLR4 and NLRP3 in the renal tissues were detected by immunohistochemical staining. The protein levels of Notch2,TLR4,NLRP3 and collagen 4( Col4) were determined by W estern blot analysis. The levels of interleukin 6( IL-6) and tum or necrosis factor( TNF-α) in the renal tissue homogenates were measured by ELISA. Spearman rank correlation test was used to analyze the correlation between Notch2,TLR4 and NLRP3 protein expression. Results Compared with the NC group,the levels of blood glucose,24-hour urine protein,kidney index,total cholesterol and triglyceride in the DM group significantly increased,while compared with the DM group,those in the ALM group were significantly reduced,but there was no significant change in blood glucose.Compared with the NC group,the protein levels of Notch2,TLR4,NLRP3 and Col4 in the renal tissues of the DM group increased. As same as the levels of Notch2,TLR4,NLRP3 and Col4 in the ALM group,the levels of IL-6 and TNF-α in the renal homogenates of the ALM group were significantly lower than those of the DM group. Correlation analysis showed that there was a significant positive correlation between the expressions of Notch2 an
作者
彭灿
蒋小涵
肖瑛
袁质平
毛彦稳
王圆圆
张莹莹
张帆
汤磊
郭兵
PENG Can;JIANG Xiaohan;XIAO Ying;YUAN Zhiping;MAO Yanwen;WANG Yuanyuan;ZHANG Yingying;ZHANG Fan;TANG Lei;GUO Bing(Department of Pathophysiology,Guizhou Medical University,Guiyang 550025,China;Key Laboratory of Drug Research and Pathogenesis of Common Chronic Diseases in Guizhou Province,Guizhou Medical University,Guiyang 550025,China;School Hospital,Guizhou Medical University,Guiyang 550025,China;Key Laboratory of Chemical Medicine,Guizhou Medical University,Guiyang 550025,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2018年第7期617-622,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
贵州省教育厅招标项目(黔教合KY字(2015)370号)
贵州省科技支撑计划(黔科合支撑[2017]2837)
贵阳市科技局基金(筑科合同[2017]30-26号)
