整合素连接激酶抑制剂在人近端小管上皮细胞转分化中的作用  

Effect of inhibitor of integrin linked kinase, QLT0267 on tubularepithelial- myofibroblast transdifferentiafion of HK- 2 Cells

作　　者：贾林[1] 林智峰[1] 马莉[1] 唐玉玲[1] 杨锐[1] 杨晓萍[1] 

机构地区：[1]新疆石河子大学医学院第一附属医院肾内科,832000

出　　处：《中华肾脏病杂志》2016年第1期50-55,共6页Chinese Journal of Nephrology

基　　金：石河子大学科学技术研究发展计划基金（2013ZRKXYQ-YD16）

摘　　要：目的观察不同浓度整合素连接激酶（ILK）抑制剂QLT0267对高糖诱导的人近端肾小管上皮细胞-肌成纤维细胞转分化（TEMT）的作用及其机制。方法体外培养人近端肾小管上皮细胞（HK-2），建立TEMT模型，排除高浓度渗透压对TEMT的影响后，将HK-2细胞分为6组，分别给予不同浓度葡萄糖（GS）及QLT0267干预48h。四甲基偶氮唑蓝（MTT）法检测细胞增殖情况，计算细胞增殖抑制率；免疫荧光法检测细胞ILK、α平滑肌肌动蛋白（d．SMA）的表达；Western印迹法检测细胞ILK、蛋白激酶B（AKT）、磷酸化蛋白激酶B（p-AKT）、α-SMA、E钙黏蛋白（E．eadherin）的表达。结果（1）与对照组相比，高糖组细胞ILK、p-AKT、α—SMA表达增高（均P〈0．05），E-cadherin表达降低（P〈0．05）。（2）与高糖组相比，浓度大于5Izmol／LQLT0267处理组的细胞增殖抑制率降低（P〈0．05）。（3）与高糖组相比，随着QLT0267浓度的增加，HK．2细胞P—AKT、ILK、α-SMA表达量逐渐降低（均P〈0．05），E—cadherin表达量逐渐增高（P〈0．05）。结论高糖可诱导HK-2细胞TEMT；ILK抑制剂QLT0267可能通过阻止ILK下游蛋白活化，部分抑制细胞增殖，延缓HK-2细胞TEMT进程。Objective To explore the effect and the possible pathway of different concentrations of QLT0267, which was the inhibitor of the integrin-linked kinase （ILK）, on the process of high glucoseinduced tubularepithelial- myofibroblast transdifferentiation （TEMT） in human renal tubular epithelial eells （HK-2）. Methods HK-2 cells were exposed to 30 mmol/L GS, and TEMT model was established. After excluding the effect of high osmotic in TEMT, HK-2 cells were divided into 6 groups by different concentrations of GS and QLT0267 for 48 hours. The rate of the cell proliferation was calculated by MTF. The expression of ILK and a-smooth muscle actin （α-SMA） were determined by immunofluoreseence and Western blot, and the expression of protein kinase B （AKT）, phosphorylated protein kinase B （p-AKT）, and E-cadherin were determined by Western blot. Results （1） The expression of ILK, p-AKT, and α-SMA in HK-2 cells were unregulated and the expression of E- cadherin was downregulated for 48 hours with glucose treating vs control （P 〈 0.05）; （2） The proliferation rate in high glucose group was higher than the group which concentration of QLT0267 was greater than 5 μmol/L （P 〈 0.05）; （3） With the concentrations of QLT0267 increased, the expression of p-AKT, α-SMA was gradually decreased （all P 〈 0.05）, and the expression of E-cadherin was gradually increased （all P 〈 0.05）. Conclusions 30 p, mol/L of GS can lead to TEMT in HK- 2 cell. TheQLT0267 with concentration greater than 5 μmol/L may prevent the activation of ILK downstream proteins, then partially inhibits cell proliferation and TEMT in HK-2 cell.

关 键 词：糖尿病肾病 细胞转分化 蛋白质丝氨酸苏氨酸激酶 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]