LncRNA-ATB在高糖诱导的人腹膜间皮细胞表型转换及增殖中的作用  

Effects of LncRNA-ATB on human peritoneal mesothelial cells phenotypic transition and proliferation induced by high glucose

作　　者：车明文 龚翔 张小金 魏达锋 王晓文 王汉民 巨立中[1] 

机构地区：[1]解放军273医院内三科 [2]武警克州支队卫生队 [3]空军广州天河干休所 [4]空军95269部队53分队保健科 [5]空军军医大学西京医院肾脏内科

出　　处：《解放军医学杂志》2017年第11期985-991,共7页Medical Journal of Chinese People's Liberation Army

摘　　要：目的探讨长链非编码RNA-ATB(Lnc RNA-ATB)在高糖刺激诱导的人腹膜间皮细胞(HPMCs)表型转换及增殖中的作用。方法将HPMCs分为对照组、甘露醇组和高糖组3组。对照组不予任何处理;高糖组和甘露醇组分别采用50mmol/L D型葡萄糖和同等渗透压的甘露醇刺激72h。采用Real-time PCR检测各组Lnc RNA-ATB、E-钙黏素(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、结缔组织生长因子(CTGF)、细胞周期素D1(Cyclin D1)、细胞周期蛋白依赖激酶抑制因子4(CDK4)、蛋白质p27(p27)及增殖细胞核抗原(PCNA)m RNA的表达,Western blotting检测各组E-cadherin、α-SMA、CTGF、Cyclin D1、CDK4、p27及PCNA蛋白的表达,并采用流式细胞仪检测细胞周期。利用慢病毒转染技术,将Lenti-Lnc RNA-ATB和sh RNA-Lnc RNA-ATB导入HPMCs中,分别上调及下调Lnc RNA-ATB的表达,采用Real-time PCR检测E-cadherin、α-SMA及CTGF m RNA的表达,Western blotting检测E-cadherin、α-SMA、CTGF蛋白的表达,流式细胞仪检测细胞周期。结果 Real-time PCR、Western blotting及流式细胞仪检测结果显示,高糖刺激后,Lnc RNA-ATB、α-SMA、CTGF、Cyclin D1、CDK4及PCNA表达增加,E-cadherin及p27表达减少(P<0.05)。上调Lnc RNAATB表达可促进细胞表型转换及增殖,下调Lnc RNA-ATB表达可抑制细胞表型转换及增殖。结论高糖刺激可通过上调Lnc RNA-ATB的表达促进人腹膜间皮细胞表型转换及增殖。Objective To explore the effect of long noncoding RNA-ATB（LncRNA-ATB） on phenotypic transition and proliferation of human peritoneal mesothelial cells（HPMCs） induced by high glucose. Methods HPMCs used in experiment were divided into three groups： control group, mannitol group and hypertonic glucose group. HPMCs in control group received no treatment, and in hypertonic glucose group and mannitol group were treated with 50 mmol/L D-glucose and isotonic mannitol for 72 hours, respectively. Real-time PCR was employed to detect the mRNA expression of LncRNA-ATB, E-cadherin, α-smooth muscle actin（α-SMA）, connective tissue growth factor（CTGF）, Cyclin D1, cyclin dependent kinase inhibitor 4（CDK4）, protein 27（p27）and proliferating cell nuclear antigen（PCNA）. Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA, CTGF, Cyclin D1, CDK4, p27 and PCNA, and flow cytometry was used to test the cell cycle. Lentivirus artifice was used to up-or down-regulate the expression of LncRNA-ATB in untreated HPMCs. Real-time PCR was employed to detect the mRNAexpression of E-cadherin, α-SMA and CTGF, Western blotting was performed to detect the proteins expression of E-cadherin, α-SMA and CTGF, and flow cytometry was used to test the cell cycle. Results It is revealed by Real-time PCR, Western blotting and flow cytometry that the expressions increased of LncRNA-ATB, α-SMA, CTGF, Cyclin D1, CDK4 and PCNA induced by hypertonic glucose, and decreased of E-cadherin and p27（P〈0.05）. Up-regulation of LncRNA-ATB promoted HPMCs phenotypic transition and proliferation, while down-regulation alleviated HPMCs phenotypic transition and proliferation. Conclusion Hypertonic glucose may accelerate HPMCs phenotypic transition and proliferation by up-regulating the expression of LncRNA-ATB.

关 键 词：细胞表型转换 LncRNA-ATB 人腹膜间皮细胞 

分 类 号：R459.51[医药卫生—治疗学]