奥美沙坦对高尿酸诱导的HK-2细胞上皮间质转化和细胞外基质合成的影响及机制  

Effects of Olmesartan on High Uric Acid-Induced Epithelial-Mesenchymal Transition and Extracellular Matrix Deposition in HK-2 Cells and Its Mechanisms

作　　者：向启蒙 耿燕秋 余燕燕 邹鑫 于冰[2] 刘秀娟[2] XIANG Qi-meng;GENG Yan-qiu;YU Yan-yan;ZOU Xin;YU Bing;LIU Xiu-juan(Graduate School of Jiangxi University of Chinese Medicine,Nanchang 330004,China;Department of Nephrology,the 908 th Hospital of PLA Joint Logistic Support Force,Nanchang 330002,China;Department of Nephrology,the Third Medical Center of PLA General Hospital,Beijing 100039,China)

机构地区：[1]江西中医药大学研究生院,南昌330004 [2]中国人民解放军联勤保障部队第九〇八医院肾内科,南昌330002 [3]中国人民解放军总医院第三医学中心肾内科,北京100039

出　　处：《南昌大学学报（医学版）》2022年第2期1-6,13,共7页Journal of Nanchang University：Medical Sciences

基　　金：国家自然科学基金(81860140);江西省自然科学基金(20181BAB205017)。

摘　　要：目的探讨奥美沙坦(OLM)对高尿酸诱导的HK-2细胞上皮间质转化和细胞外基质合成的影响及机制。方法体外培养的HK-2细胞分为5组:对照组(正常培养)、高尿酸组(600μmol·L^(-1)尿酸处理)、高尿酸+OLM组(高尿酸联合OLM处理)、高尿酸+OLM+Toll样受体4(TLR4)组(TLR4过表达质粒转染细胞用高尿酸联合OLM处理)、高尿酸+OLM+空载组(空载质粒转染细胞用高尿酸联合OLM处理)。CCK-8法检测各组细胞活力,western blot法检测上皮细胞标志蛋白E-cadherin和间质细胞标志蛋白α-SMA、vimentin表达,ELISA法检测细胞外基质(collagenⅠ和Fibronectin)、炎症因子(IL-6和TNF-α)水平,试剂盒检测活性氧ROS水平。结果与对照组比较,高尿酸组HK-2细胞的E-cadherin蛋白表达降低(P<0.05),vimentin、α-SMA、TLR4蛋白表达增加(均P<0.05),collagenⅠ、Fibronectin、IL-6、TNF-α和ROS的水平均增加(均P<0.05);与高尿酸组比较,高尿酸+OLM组细胞E-cadherin蛋白表达增加(P<0.05),而vimentin、α-SMA、TLR4蛋白表达降低(均P<0.05),collagenⅠ、Fibronectin、IL-6、TNF-α和ROS的水平降低(均P<0.05);与高尿酸+OLM组比较,高尿酸+OLM+TLR4组TLR4、vimentin、α-SMA蛋白表达显著增加(均P<0.05),E-cadherin蛋白表达降低(P<0.05),collagenⅠ、Fibronectin、IL-6、TNF-α和ROS的水平增加(均P<0.05)。结论OLM可抑制高尿酸诱导的HK-2细胞上皮间质转化和细胞外基质合成,其机制是通过抑制TLR4及其介导的氧化应激和炎症反应而实现。Objective To investigate the effects of olmesartan(OLM)on high uric acid-induced epithelial-mesenchymal transition and extracellular matrix deposition in HK-2 cells and its mechanisms.Methods The HK-2 cells were divided into five groups:control group(cells were cultured under normal condition),high uric acid group(cells were treated with 600μmol·L^(-1) uric acid alone),high uric acid+OLM group(cells were treated with uric acid and OLM),high uric acid+OLM+TLR4 group(cells were transfected with TLR4 overexpression plasmids and treated with uric acid and OLM),and high uric acid+OLM+empty vector group(cells were transfected with empty vectors and treated with uric acid and OLM).Cell viability was detected by CCK-8 assay.The expression of epithelium marker E-cadherin and mesenchymal markerα-SMA and vimentin was detected by Western blot.The levels of extracellular matrix collagenⅠand fibronectin and inflammatory factors interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were measured by ELISA.The production of reactive oxygen species(ROS)was determined by a commercial kit.Results Compared with the control group,treatment with uric acid alone decreased the expression of E-cadherin,but increased the expression of vimentin,α-SMA and TLR4 and the levels of collagenⅠ,fibronectin,IL-6,TNF-αand ROS(P<0.05).Compared with high uric acid group,E-cadherin expression increased but vimentin,α-SMA and TLR4 expression and collagenⅠ,fibronectin,IL-6,TNF-αand ROS levels decreased in high uric acid+OLM group(P<0.05).Compared with high uric acid+OLM group,the expression of TLR4,vimentin andα-SMA and the levels of collagenⅠ,fibronectin,IL-6,TNF-αand ROS increased but the expression of E-cadherin decreased in high uric acid+OLM+TLR4 group(P<0.05).Conclusion OLM treatment can suppress high uric acid-induced epithelial-mesenchymal transition and extracellular matrix deposition in HK-2 cells through inhibiting TLR4-mediated oxidative stress and inflammatory response.

关 键 词：奥美沙坦 肾纤维化 Toll样受体4 上皮间质转化 细胞外基质 

分 类 号：R692[医药卫生—泌尿科学] R965[医药卫生—外科学]