高糖对大鼠腹膜间皮细胞整合素连接激酶、结缔组织生长因子及α-平滑肌肌动蛋白表达的影响  

The expression of connective tissue growth factor, integrin linked kinase and SMA in rat peritoneal mesothelial cells induced by high glucose

作　　者：孙立霞[1] 马健飞[1] 邓文艳[1] 

机构地区：[1]中国医科大学附属第一医院肾脏内科,沈阳110001

出　　处：《中国血液净化》2013年第6期323-326,共4页Chinese Journal of Blood Purification

摘　　要：目的研究高糖作用下大鼠腹膜间皮细胞(rat peritoneal mesothelial cells,RPMC)整合素连接激酶(integrin liked kinase,ILK)、结缔组织生长因子(connective tissue growth factor,CTGF)及α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达的情况,为防治腹膜透析相关性腹膜纤维化提供新的思路。方法胰蛋白酶消化法原代及传代培养RPMC,经鉴定后第2代用于实验。细胞随机分组:①正常对照组;②高糖组:不同浓度的高糖(1.5%、2.5%、4.25%)作用24h;2.5%高糖分别作用RPMC12h、24h、48h、72h。实时定量PCR检测ILK及α-SMAmRNA的表达。Western印迹法检测ILK蛋白的表达。ELISA法检测细胞培养上清液中CTGF的表达。结果与正常对照组相比,高糖可刺激RPMCILK及α-SMA的表达增高(均P<0.05),高糖诱导RPMCCTGF的表达增高(P<0.05)。结论高糖可上调RPMCILK、CTGF及α-SMA的表达。ILK及CTGF参与了高糖诱导的大鼠腹膜间皮细胞腹膜纤维化过程,为防治腹膜透析相关性腹膜纤维化提供新的思路。Objective To observe the expression of integrin linked kinase （ILK）, connective tissue growth factor （CTGF） and α-smooth muscle actin （a-SMA） in rat peritoneal mesothelial cells （RPMCs） in- duced by high glucose. Methods RPMCs were isolated, cultured and amplified by trypsin, then identified. RPMCs of second passage were used for the experiment. RPMCs were divided into normal control group, high glucose （1.5%, 2.5%, and 4.25%） for 24 hours groups, and high glucose （2.5%） for 12, 24, 48, and 72 hours groups. ILK and α-SMA mRNAs were detected by real time RT-PCR. ILK protein was detected by western blot. CTGF protein in supematant was detected by ELISA. Results Compared with the control group, the expression of ILK, CTGF and α-SMA were significantly increased in the groups stimulated by high glucose （P 〈 0.05）. Conclusions High glucose can up-regulate the expression of ILK, CTGF and α-SMA in RPMCs. ILK and CTGF may play a role in the pathogenesis of peritoneal dialysis related peritoneal fibrosis.

关 键 词：腹膜间皮细胞 高糖 整合素连接激酶 结缔组织生长因子 纤维化 

分 类 号：R459.5[医药卫生—治疗学]