Yi Bin, Hu Xiaofang, Zhang Hao, Huang Jing, Liu Jishi, Hu Jing, Li Wei, Huang Lihua
Department of Nephrology, The Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
Medical experimental center, The Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
PLoS One. 2014 Jun 17;9(6):e99633. doi: 10.1371/journal.pone.0099633. eCollection 2014.
To investigate if nuclear NF-κB p65 expression in ex vivo isolated peripheral blood mononuclear cells correlates with urinary MCP-1 or RANTES and the severity of type 2 diabetic nephropathy.
According to their urinary albumin-to-creatinine ratio (uACR), 107 patients with type 2 diabetes (eGFR >60 ml/min) were divided into normal albuminuria group (DN0 group, 38 cases), microalbuminuria group (DN1 group, 38 cases), and macroalbuminuria group (DN2 group, 31 cases), compared with matched healthy normal control group (NC group, 30 cases). Nuclear NF-κB p65 protein expression levels in peripheral blood mononuclear cells were detected by western blotting. Real-time quantitative polymerase chain reaction was used to detect NF-κB p65 mRNA expression and ELISA assay was used to detect the levels of urinary MCP-1 and RANTES.
Nuclear NF-κB p65 protein and NF-κB p65 mRNA expression levels in peripheral blood mononuclear cells, urinary MCP-1/Cr and RANTES/Cr were all significantly higher in all diabetes groups as compared with NC group. In particular, the increase of nuclear NF-κB p65 protein and NF-κB p65 mRNA expressions, urinary MCP-1/Cr and RANTES/Cr all correlated with the severity of type 2 diabetic nephropathy as indicated by the increase in uACR. Pearson correlation analysis indicated that both urinary MCP-1/Cr and RANTES/Cr were positively correlated with nuclear NF-κB p65 protein or NF-κB p65 mRNA levels. Stepwise multiple regression analysis showed that nuclear NF-κB p65 protein or NF-κB p65 mRNA was an independent variable for urinary MCP-1/Cr, and MCP-1/Cr and RANTES/Cr were two independent variables for uACR.
Our research demonstrates that nuclear NF-κB p65 protein and mRNA expressions in ex vivo isolated peripheral blood mononuclear cells well correlate with urinary MCP-1/Cr, RANTES/Cr and the severity of type 2 diabetic nephropathy.
研究体外分离的外周血单个核细胞中核因子κB p65（NF-κB p65）的表达与尿单核细胞趋化蛋白-1（MCP-1）或调节激活正常T细胞表达和分泌因子（RANTES）以及2型糖尿病肾病严重程度之间的相关性。
根据尿白蛋白与肌酐比值（uACR），将107例2型糖尿病患者（估算肾小球滤过率[eGFR]>60 ml/min）分为正常蛋白尿组（DN0组，38例）、微量白蛋白尿组（DN1组，38例）和大量白蛋白尿组（DN2组，31例），并与年龄匹配的健康正常对照组（NC组，30例）进行比较。采用蛋白质免疫印迹法检测外周血单个核细胞中核NF-κB p65蛋白表达水平。采用实时定量聚合酶链反应检测NF-κB p65 mRNA表达水平，酶联免疫吸附测定法检测尿MCP-1和RANTES水平。
与NC组相比，各糖尿病组外周血单个核细胞中核NF-κB p65蛋白和NF-κB p65 mRNA表达水平、尿MCP-1/Cr和RANTES/Cr均显著升高。特别是，核NF-κB p65蛋白和NF-κB p65 mRNA表达的增加、尿MCP-1/Cr和RANTES/Cr均与uACR升高所提示的2型糖尿病肾病严重程度相关。Pearson相关分析表明，尿MCP-1/Cr和RANTES/Cr均与核NF-κB p65蛋白或NF-κB p65 mRNA水平呈正相关。逐步多元回归分析显示，核NF-κB p65蛋白或NF-κB p65 mRNA是尿MCP-1/Cr的独立变量，而MCP-1/Cr和RANTES/Cr是uACR的两个独立变量。
我们的研究表明，体外分离的外周血单个核细胞中核NF-κB p65蛋白和mRNA表达与尿MCP-1/Cr、RANTES/Cr以及2型糖尿病肾病严重程度密切相关。
