| 刘立栋,徐腊梅,高洁,等.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[J].安徽医药,2023,27(1):13-18. |
| 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 |
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| DOI：10.3969/j.issn.1009-6469.2023.01.004 |
| 中文关键词: 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 |
| 英文关键词: Hyperglycemia Diabetic nephropathy Resveratrol 微小 RNA-574-3p Glomerular mesangial cells Inflammacript-tion Oxidative stress Rats |
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| 中文摘要: |
| 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 +5 μmoL白藜芦醇)、 HG+中剂量组( 30 mmoL葡萄糖 +10 μmoL白藜芦醇)、 HG+高剂量组( 30 mmoL葡萄糖 +20 μmoL白藜芦醇)、 HG+中剂量 +anti-miR-NC组( 30 mmoL葡萄糖 +10 μmoL白藜芦醇 +转染 anti-miR-NC)、 HG+中剂量 +anti-miR-574-3p组( 30 mmoL葡萄糖 +10 μmoL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)、 Cyclin 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、细胞迁移数、 IL-1β(63.33±1.15比 41.78±1.79)μg/L、TNF-α、丙二醛( 51.56±1.52比 35.89±1.28)μmoL/g、LDH水平均升高， p21蛋白表达量降低，差异有统计学意义( P<0.05)。结论白藜芦醇可能通过上调高糖诱导的大鼠肾小球系膜细胞中 miR574-3p表达，抑制细胞增殖、迁移，并减轻其炎症水平和氧化应激。 |
| 英文摘要: |
| Objective To investigate whether resveratrol affects the proliferation, inflammation, and oxidative stress of diabetic nephropathy 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+5 μmoL resveratrol), HG+medium-dose group (30 mmoL glucose+10 μmoL resveratrol), HG+high-dose group (30 mmoL glucose+20 μmoL resveratrol), HG+medium dose+anti-miR-NC group (30 mmoL glucose+10 μmoL resveratrol+transfected anti-miR-NC), HG+medium dose+anti-miR-574-3p group (30 mmoL glucose+10 μmoL 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 immunosorbent assay (ELISA) and test kits were performed to detect the proliferation of RMC, expression of Cyclin D1 (Cyclin D1), expression ofp21, 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-dose group, and HG+high-dose group, while the expression of p21 protein, miR-574-3p [(0.48±0.02) vs. (0.27±0.02), (0.65±0.02) vs. (0.27±0.02), (0.83±0.02) vs. (0.27±0.02)] were increased, and the differences were statistically significant (P<0.05). Compared with the HG+medium dose group, the cell activity [(1.91±0.02) vs. (1.66± 0.03)] of RMC, the protein expression of Cyclin D1, cell migration number, levels of IL-1β [(63.33±1.15) vs. (41.78±1.79) μg/L], TNFα, MDA [(51.56±1.52) vs. (35.89±1.28) μmoL/g], and LDH in the HG+medium dose+anti-miR-574-3p group were increased, but the expression of p21 protein was decreased, and the differences were statistically significant (P<0.05).Conclusion Resveratrol may inhibit cell proliferation, migration, the level of inflammation, and oxidative stress by up-regulating the expression of miR-574-3p in high glucose-induced rat glomerular mesangial cells. |
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