lncRNA PACER对高糖诱导的人肾小管上皮细胞中炎性因子表达和转分化的影响  

Effects of lncRNA PACER on the Expressions of Inflammatory Factors and Transdifferentiation in Human Renal Tubular Epithelial Cells Induced by High Glucose

作　　者：李海燕 杨彦平 刘国 Li Haiyan;Yang Yanping;Liu Cuo(Department of Geriatrics,No.215 Hospital of Shaanxi Nuclear Industry,Shaanxi Xianyang 712000,China;Department of Nephrology,No.215 Hospital of Shaanxi Nuclear Industry,Shaanxi Xianyang 712000,China)

机构地区：[1]陕西省核工业二一五医院老年病科,陕西咸阳712000 [2]陕西省核工业二一五医院肾内科,陕西咸阳712000

出　　处：《中国药师》2021年第9期1647-1651,共5页China Pharmacist

摘　　要：目的:探讨P50相关的环氧合酶2外源性RNA(PACER)对高糖诱导的人肾小管上皮细胞中炎性因子表达和转分化的影响。方法:实时荧光定量PCR(RT-qPCR)检测60例单纯2型糖尿病(T2D)患者(T2D组)、60例糖尿病肾病(DN)患者(DN组)及60例健康体检者(正常对照组)血清中PACER表达水平。体外培养人肾小管上皮细胞HK-2,转染PACER小干扰RNA后用25 mmol·L-1葡萄糖诱导24 h,RT-qPCR检测细胞中PACER表达水平,酶联免疫吸附法检测细胞培养上清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和单核细胞趋化因子-1(MCP-1)表达,蛋白印迹(Western Blot)法检测细胞中E-钙黏蛋白(E-cadherin)、α平滑肌肌动蛋白(α-SMA)、核因子-κB p65(NF-κB p65)和磷酸化的NF-κB抑制蛋白α(p-IκBα)。结果:与正常对照组比较,T2D组和DN组患者血清中PACER表达水平升高(P<0.05);与T2D组比较,DN组患者血清中PACER表达水平升高(P<0.05)。高糖作用HK-2细胞后,HK-2细胞中PACER表达水平升高(P<0.05),细胞培养上清液中TNF-α、IL-6和MCP-1表达水平均升高(P<0.05),细胞中E-cadherin蛋白表达水平降低(P<0.05),α-SMA、NF-κB p65和p-IκBα蛋白表达水平均升高(P<0.05)。下调PACER降低了高糖诱导的HK-2细胞培养上清液中TNF-α、IL-6和MCP-1的表达及细胞中α-SMA、NF-κB p65和p-IκBα的蛋白表达(P<0.05),促进了细胞中E-cadherin蛋白表达(P<0.05)。结论:PACER在DN患者血清和高糖诱导的HK-2细胞中表达升高,下调PACER表达可有效抑制高糖诱导的HK-2细胞中炎性因子的表达及EMT过程,其作用机制可能与NF-κB信号通路有关。Objective:To investigate the effects of P50-related cyclooxygenase 2 exogenous RNA(PACER)on the expressions of inflammatory factors and transdifferentiation in human renal tubular epithelial cells induced by high glucose.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression of PACER in serum of 60 patients with type 2 diabetes mellitus(T2 D group),60 patients with T2 D nephropathy(DN group)and 60 healthy people(normal control group).Human renal tubular epithelial cells HK-2 were cultured in vitro and transfected with PACER small interfering RNA,and then they were induced with 25 mmol·L-1 glucose for 24 h.RT-qPCR was used to detect the expression level of PACER,enzyme-linked immunosorbent assay was used to detect the expression levels of TNF-α,IL-6 and MCP-1 in cell culture supernatants,and Western blot method was used to detect the expression levels of E-cadherin,α-SMA,NF-κB p65 and p-IκBαprotein.Results:Compared with those in the normal control group,the expression level of PACER in serum of patients in T2 D group and DN group increased(P<0.05).Compared with those of T2 D group,the expression level of PACER in serum of patients with DN group increased(P<0.05).After high glucose induced HK-2 cells,the expression level of PACER in HK-2 cells increased(P<0.05),the expression levels of TNF-α,IL-6 and MCP-1 in cell culture supernatants increased(P<0.05),the expression level of E-cadherin protein was significantly reduced in cells(P<0.05),and the expression levels ofα-SMA,NF-κB p65 and p-IκBαprotein increased(P<0.05).Down-regulating PACER reduced the levels of TNF-α,IL-6 and MCP-1 in the culture supernatant of HK-2 cells induced by high glucose and the protein expressions ofα-SMA,NF-κB p65 and p-IκBαin HK-2 cells induced by high glucose(P<0.05),while increased the protein expression of E-cadherin(P<0.05).Conclusion:The expression of PACER is increased in serum of DN patients and high glucose-induced HK-2 cells.Down-regulating the expression of PACER can effectively i

关 键 词：肾小管上皮细胞 P50相关的环氧合酶2外源性RNA 高糖 炎性因子 转分化 

分 类 号：R965.1[医药卫生—药理学]