目的 探讨Crk1/2与CrkL缺失导致足细胞损伤过程中胞内蛋白表达变化情况.方法 利用小干扰RNA转染方法对足细胞内Crk1/2与CrkL进行单敲降及双敲降;通过非标记液相色谱串联质谱方法对Crk1/2与CrkL单敲降及双敲降的足细胞进行蛋白质组学分析;对得到的差异蛋白进行基因本体(GO)分析和京都基因和基因组百科全书(KEGG)分析;采用Western blot试验对差异蛋白进行验证.结果 Crk1/2与CrkL单敲降及双敲降的足细胞与正常足细胞比较,共发现98个差异蛋白,GO分析和KEGG分析提示多数上调蛋白富集在代谢途径,多数下调蛋白富集在信号转导途径,如抑制和激活蛋白1、磷脂酰肌醇三激酶-丝氨酸/苏氨酸激酶和环磷酸腺苷信号途径.经Western blot试验验证,细胞代谢相关蛋白超氧化物歧化酶2(SOD2)表达上调,信号转导相关蛋白L RP1表达上调、c-Jun表达下调,细胞骨架损伤相关蛋白T PM 4表达上调、Cdc42EP1表达下调.结论 Crk1/2与CrkL缺失可通过调节代谢途径及相关信号转导途径导致足细胞损伤,TPM4、SOD2、LRP1、c-Jun和Cdc42EP1参与损伤过程,具体分子机制值得进一步探讨.

目的 探讨人近端肾小管上皮细胞(HK-2细胞系)在糖化白蛋白(glycated albumin,GA)与齐墩果酸(oleanolic acid,OA)作用下,肾损伤分子-1(kidney injury molecule-1,KIM-1)、中性粒细胞明胶酶相关载脂蛋白(neutrophil gelatinase-associated lipocalin,NGAL)及Toll样受体信号通路相关分子的表达情况.方法 采用GA及OA处理HK-2细胞24h,real-time PCR检测KIM-1、NGAL以及p38丝裂原活化蛋白激酶(mitogen activated protein kinase,MAPK)、白细胞介素-1受体相关激酶4(interleukin-1 receptor-associated kinase 4,IRAK4)、Toll样受体(Toll-like receptor,TLR)1、TLR2、TLR7和TLR9的mRNA表达,ELISA法检测细胞上清液KIM-1与NGAL水平,Western blotting法检测细胞内p38 MAPK、IRAK4、TLR1、TLR2、TLR7和TLR9蛋白表达.结果 与对照组比较,GA能显著上调KIM-1、NGAL mRNA与蛋白的含量(P＜0.05),并促进p38 MAPK、IRAK4、TLR1、TLR2 mRNA及蛋白的表达(P＜0.05);加入OA能显著抑制上述作用(P＜0.05).结论 GA能够上调KIM-1和NGAL的表达与释放,激活Toll样受体信号通路,对人近端肾小管上皮细胞造成损伤;而加入OA能够有效干预GA调控的损伤.

The complement system is a complex protein network.Complement is important in disease severity and response to therapy.This review summarizes recent advances on the role of complement analysis in complement function or activity,complement factors concentrations,complement activation products,complement autoantibody and genetic analysis.And it also focuses on pre-analytical and post-analytical quality control for complement analysis.

Diabetic kidney disease is one of the main causes of end-stage renal disease .During the past decades , proteomic analysis based on mass spectrometry has been playing a momentous role in revealing potential biomarkers of diabetic kidney disease .Assays using New combination of multiple biomarkers can help identify high risk patients and give positive guidance of therapies .