整合素连接激酶抑制剂对人肾小管上皮细胞转分化的影响  

Influence of integrin-linked kinase inhibitor on renal tubular epithelial-mesenchymal transdifferentiation

作　　者：林智峰[1] 贾林[1] 张春江[1] 杨晓萍[1] LIN Zhi-feng;JIA Lin;ZHANG Chun-jiang;YANG Xiao ping(Department of Nephrolog y , the First Affiliated Hospital of Shihezi University, Shihezi 832008, Chin)

机构地区：[1]石河子大学医学院第一附属医院肾病科,新疆石河子832008

出　　处：《中华实用诊断与治疗杂志》2018年第7期646-649,共4页Journal of Chinese Practical Diagnosis and Therapy

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

摘　　要：目的探讨整合素连接激酶(integrin-linked kinase,ILK)抑制剂在人近端肾小管上皮细胞转分化中的作用及可能机制。方法对数生长期人近端肾小管上皮细胞(HK-2)随机分为对照组(无糖DMEM/F12培养液)、高糖组(30.0mmol/L葡萄糖DMEM/F12培养液)、抑制剂组(30.0 mmol/L葡萄糖DMEM/F12培养液+30.0μmol/L QLT0267 100μL),培养48h后,倒置相差显微镜下观察细胞形态,Western blot法检测HK-2细胞ILK、E-钙黏蛋白(E-cadherin)、蛋白激酶B(protein kinase B,Akt)、磷酸化Akt(phosphorylated Akt,P-Akt)相对表达量,反转录PCR法检测ILK mRNA、纤连蛋白(fibronectin,Fn)mRNA表达量。结果对照组HK-2细胞呈椭圆形或圆形,细胞间连接紧密,呈岛屿状生长;高糖组细胞向长梭型转变,细胞间紧密连接消失,呈离散状生长;抑制剂组细胞改变同高糖组,但程度较轻;高糖组、抑制剂组HK-2细胞E-cadherin相对表达量(0.43±0.04、0.80±0.05)均低于对照组(1.04±0.05)(P<0.05),且高糖组低于抑制剂组(P<0.05);高糖组、抑制剂组P-Akt相对表达量(0.49±0.05、0.31±0.03)均高于对照组(0.21±0.01)(P<0.05);且高糖组高于抑制剂组(P<0.05),高糖组、抑制剂组HK-2细胞ILK相对表达量(0.52±0.02、0.51±0.02)均高于对照组(0.22±0.01)(P<0.05),高糖组与抑制剂组比较差异无统计学意义(P>0.05);高糖组、抑制剂组及对照组HK-2细胞Akt相对表达量(0.91±0.03、0.92±0.02、0.94±0.05)比较差异均无统计学意义(P>0.05);高糖组、抑制剂组HK-2细胞ILK mRNA(1.83±0.23、1.64±0.07)、Fn mRNA相对表达量(2.06±0.07、1.79±0.06)均高于对照组(0.83±0.04、1.14±0.08)(P<0.05),且高糖组Fn mRNA相对表达量高于抑制剂组(P<0.05),ILK mRNA相对表达量与抑制剂组比较差异无统计学意义(P>0.05)。结论ILK抑制剂QLT0267可抑制ILK下游效应分子Akt的磷酸化,减少Fn、E-cadherin下调,从而抑制或延缓肾小管上皮细胞转分化进程。Objective To explore the role and mechanism of integrin-linked kinase（ILK）inhibitors in the transdifferentiation of proximal renal tubular epithelial cells.Methods The proximal renal tubular epithelial cells in logarithmic phase（HK-2）were randomly divided into control group（glucose-free DMEM/F12 culture）,and high-glucose group（30.0 mmol/L glucose DMEM/F12 culture）,and inhibitor group（30.0 mmol/L glucose DMEM/F12 culture ＋30.0μmol/L QLT0267 100μL）.After culture for 48 h,the cellular morphology was observed under inverted phase contrast microscope.Western blot method was used to detect the relative expressions of ILK,E-cadherin,protein kinase B（Akt）and phosphorylated Akt（P-Akt）in HK-2 cells,and the expressions of ILK mRNA and fibronectin（Fn）mRNA were detected by reverse transcription PCR.Results The HK-2 cells showed round or oval shape,compact connection between cells and island shaped growth in control group.The HK-2 cells showed changing to long spindle shape,disappeared close connection between cell,and discrete shaped growth in high-glucose group.The changes of HK-2 cells in inhibitor group were milder than those in high-glucose group.The relative expression of E-cadherin was significantly lower in high-glucose group（0.43±0.04）and inhibitor group（0.80±0.05）than that in control group（1.04±0.05）（P〈0.05）,and in high-glucose group than that in inhibitor group（P〈0.05）.The relative expression of P-Akt was significantly higher in high-glucose group（0.49±0.05）and inhibitor group（0.31±0.03）than that in control group（0.21±0.01）（P〈0.05）,and in high-glucose group than that in inhibitor group（P〈0.05）.The relative expression of ILK was significantly higher in high-glucose group（0.52±0.02）and inhibitor group（0.51±0.02）than that in control group（0.22±0.01）（P〈0.05）,and there was no significant difference between high-glucose group and inhibitor group（P〉0.05）.There was no significant difference in the Akt relativ

关 键 词：人肾小管上皮细胞 上皮细胞转分化 整合素链接激酶 QLT0267 高糖 

分 类 号：R692[医药卫生—泌尿科学]