CXCL16基因沉默减轻ox-LDL对小鼠足细胞损伤  

CXCL16 gene silencing alleviates injury of mouse podocytes treated with the oxidized low-density lipoprotein

作　　者：王雪[1] 李倩[1] 王莉[1] 孙书珍[1] 马爱华[2] 

机构地区：[1]山东大学附属省立医院小儿肾脏风湿免疫科,山东济南250021 [2]山东大学附属省立医院小儿神经科,山东济南250021

出　　处：《山东大学学报（医学版）》2016年第6期16-21,共6页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(ZR2010HM110);山东省自然科学基金(ZR2015HM009);山东省科技发展计划(2014GGH218009)

摘　　要：目的探讨CXC趋化因子配体16(CXCL16)基因沉默在减轻氧化低密度脂蛋白(ox-LDL)对足细胞损伤中的作用。方法小鼠CXCL16基因短发卡RNA(shRNA)慢病毒及阴性对照慢病毒转染足细胞,Real-time PCR、Western blotting检测转染效率。CCK8法检测转染后足细胞活力。将转染后足细胞分为4组培养和处理,即阴性对照组(NC组)、ox-LDL组、CXCL16-shRNA组、ox-LDL+CXCL16-shRNA组。总胆固醇含量测定估计细胞内脂质沉积,Western blotting检测CXCL16及β1整合素变化,并检测活性氧(ROS)的表达。结果 CXCL16-shRNA慢病毒转染后,CXCL16 mRNA抑制率约70%(P=0.008),CXCL16蛋白抑制率约60%(P=0.008)。CXCL16-shRNA组及NC组细胞活力差异无统计学意义(P=0.712)。ox-LDL+CXCL16-shRNA组较ox-LDL组细胞内脂质沉积减少(P<0.001),β1整合素表达量增加(P=0.004),ROS产生减少(P<0.001)。结论 CXCL16基因沉默可减少ox-LDL作用后ROS表达量,并增加β1整合素的表达,发挥足细胞保护作用。Objective To investigate the role of chemokine ligand 16（ CXCL16） gene silencing on alleviating injury of podocytes treated with oxidized low-density lipoprotein（ ox-LDL）. Methods The mouse podocytes were transfected with CXCL16 gene short hairpin RNA（ shRNA） lentivirus vector and negative control lentivirus vector. The protein and mRNA expressions of CXCL16 were determined with Western blotting and Real-time PCR. The vitality of podocytes was detected with CCK8 assay. The mouse podocytes（ cell line） were divided into 4 groups： negative control group（ NC group）,ox-LDL group,CXCL16-shRNA group,and ox-LDL ＋ CXCL16-shRNA group. The total cholesterol was detected with cholorimetric detection to measure intracelluar lipid accumulations. The expressions of CXCL16 and integrin-β1 were detected with Western blotting. The expression of ROS was detected with ROS Deep Red Dye.Results The inhibition rate of CXCL16 mRNA was about 70%,and that of CXCL16 protein was about 60%,compared to the negative control（ both P = 0. 008）. CCK8 revealed no significant difference in the vitality of podocytes between NC group and CXCL16-shRNA group（ P = 0. 712）. Compared to the ox-LDL group,in the ox-LDL ＋ CXCL16-shRNA group,the intracelluar lipid accumulations were reduced（ P〈0. 001）,integrin-β1 was increased（ P = 0. 004）,and ROS was reduced（ P〈0. 001）. Conclusion CXCL16 gene silencing is likely to play an important role for podocytic protection after the treatment of ox-LDL by reducing the synthesis of ROS and elevating the expression of integrin-β1.

关 键 词：足细胞 CXC趋化因子配体16 短发卡RNA 氧化低密度脂蛋白 活性氧 Β1整合素 

分 类 号：R726.9[医药卫生—儿科]