白藜芦醇通过调控微小RNA-574-3p对高糖环境下肾小球系膜细胞增殖、炎症水平及氧化应激的影响研究  

Effect of resveratrol on cell proliferation,inflammation,and oxidative stress by regulating miR-574-3p in high glucose-induced glomerular mesangial cells

作　　者：刘立栋 徐腊梅 高洁 李洁 邵英 田雄涛 刘晓宇 LIU Lidong;XU Lamei;GAO Jie;LI Jie;SHAO Ying;TIAN Xiongtao;LIU Xiaoyu(Department of Endocrinology and Metabolism,986 Hospital,PLA Air Force,Xi'an,Shaanxi 710000,China)

机构地区：[1]中国人民解放军空军第九八六医院内分泌科,陕西西安710000

出　　处：《安徽医药》2023年第1期13-18,I0003,共7页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨白藜芦醇是否通过微小RNA(miRNA)-574-3p影响糖尿病肾病细胞的增殖、炎症水平及氧化应激。方法在2019年1月1日至2020年6月1日,将大鼠肾小球系膜细胞(RMC)分为NG组(5.3 mmoL葡萄糖)、HG组(30 mmoL葡萄糖)、HG+低剂量组(30 mmoL葡萄糖+5µmoL白藜芦醇)、HG+中剂量组(30 mmoL葡萄糖+10µmoL白藜芦醇)、HG+高剂量组(30 mmoL葡萄糖+20µmoL白藜芦醇)、HG+中剂量+anti-miR-NC组(30 mmoL葡萄糖+10µmoL白藜芦醇+转染anti-miR-NC)、HG+中剂量+anti-miR-574-3p组(30 mmoL葡萄糖+10µmoL白藜芦醇+转染anti-miR-574-3p)。应用噻唑基蓝四唑溴化物(MTT)法、蛋白质印迹法(western blotting)、Transwell法、实时定量聚合酶链反应(qRT-PCR)、酶联免疫吸附测定(ELISA)、试剂盒分别检测RMC增殖、细胞周期蛋白D1(Cyclin D1)表达、p21表达、细胞迁移、miR-574-3p表达、白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)、丙二醛、乳酸脱氢酶(LDH)水平。结果与NG组相比,HG组RMC的细胞活性(1.97±0.039比1.39±0.02)、Cyclin D1蛋白表达量、细胞迁移数、IL-1β(70.44±1.74比20.67±0.78)µg/L、TNF-α、丙二醛(49.22±1.06比13.33±0.67)µmoL/g、LDH水平升高,p21蛋白表达量、miR-574-3p表达量降低(0.27±0.02比1.04±0.02),差异有统计学意义(P<0.05)。与HG组相比,HG+低剂量组、HG+中剂量组和HG+高剂量组RMC细胞活性(1.74±0.02比1.97±0.03,1.64±0.02比1.97±0.03,1.53±0.01比1.97±0.03)、Cy‐clin D1蛋白表达量、细胞迁移数、IL-1β(53.33±1.53比70.44±1.74,40.78±1.34比70.44±1.74,28.33±1.20比70.44±1.74)µg/L、TNF-α、丙二醛(41.22±0.86比49.22±1.06,30.78±0.91比49.22±1.06,20.89±0.71比49.22±1.06)µmoL/g、LDH水平均降低,p21蛋白表达量、miR-574-3p表达量(0.48±0.02比0.27±0.02,0.65±0.01比0.27±0.02,0.83±0.02比0.27±0.02)升高,差异有统计学意义(P<0.05)。与HG+中剂量组相比,HG+中剂量+anti-miR-574-3p组RMC的细胞活性(1.91±0.02比1.66±0.03)、Cyclin D1蛋白表达量�Objective To investigate whether resveratrol affects the proliferation,inflammation,and oxidative stress of diabetic ne‐phropathy cells through microRNA(miRNA)-574-3p.Methods Between January 1,2019 and June 1,2020,rat mesangial cells(RMC)were assigned into NG group(5.3 mmoL glucose),HG group(30 mmoL glucose),HG+low-dose group(30 mmoL glucose+5µmoL resve‐ratrol),HG+medium-dose group(30 mmoL glucose+10µmoL resveratrol),HG+high-dose group(30 mmoL glucose+20µmoL resvera‐trol),HG+medium dose+anti-miR-NC group(30 mmoL glucose+10µmoL resveratrol+transfected anti-miR-NC),HG+medium dose+an‐ti-miR-574-3p group(30 mmoL glucose+10µmoL resveratrol+transfected anti-miR-574-3p).Thiazolyl blue tetrazolium bromide(MTT)method,Western blotting,Transwell method,real-time quantitative polymerase chain reaction(qRT-PCR),enzyme-linked immunosor‐bent assay(ELISA)and test kits were performed to detect the proliferation of RMC,expression of Cyclin D1(Cyclin D1),expression of p21,cell migration,expression of miR-574-3p,and levels of interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),malondialdehyde(MDA),and lactate dehydrogenase(LDH).Results Compared with the NG group,the cell activity[(1.97±0.039)vs.(1.39±0.03)]of RMC,the protein expression of Cyclin D1,cell migration number,levels of IL-1β[70.44±1.74)vs.(20.67±0.78)µg/L],TNF-α,MDA[(49.22±1.06)vs.(13.33±0.67)µmoL/g],and LDH were increased in the HG group,the expressions of p21 protein and miR-574-3p[(0.27±0.02)vs.(1.04±0.02)]were decreased,and the differences were statistically significant(P<0.05).Compared with the HG group,the cell activity[(1.74±0.02)vs.(1.97±0.03),(1.64±0.02)vs.(1.97±0.03),(1.53±0.01)vs.(1.97±0.03)]of RMC,the protein expression of Cyclin D1,cell migration number,levels of IL-1β[(53.33±1.53)vs.(70.44±1.53),(40.78±1.53 vs.(70.44±1.53),(28.33±1.53)vs.(70.44±1.74)µg/L],TNF-α,MDA[(41.22±0.86)vs.(49.22±0.86),(30.78±0.86)vs.(49.22±0.86),(20.89±0.86)vs.(49.22±1.06)µmoL/g],and LDH were reduced in the HG+low-dose group,HG+medium

关 键 词：高血糖症 糖尿病肾病 白藜芦醇 miR-574-3p 肾小球系膜细胞 炎症 氧化应激 大鼠 

分 类 号：R285[医药卫生—中药学]