Department of Nephrology, First Hospital of China Medical University, Shenyang, Liaoning, China (mainland).
Med Sci Monit. 2018 Feb 11;24:846-854. doi: 10.12659/msm.907814.
BACKGROUND To investigate the protective effect of ursolic acid (UA) on high glucose (HG)-induced human glomerular mesangial cell injury and to determine whether UA inhibits cell proliferation and reactive oxygen species (ROS) production by suppressing PI3K/Akt/mTOR pathway activation. MATERIAL AND METHODS Human mesangial cells were cultured with normal glucose (NG group), high glucose (HG group), mannitol (mannitol hypertonic control group), or high glucose with different concentrations (0.5, 1.0, and 2.0 mmol/L) of UA (HG+UA groups). Cell proliferation and intracellular ROS levels were assessed by methyl thiazolyl tetrazolium (MTT) and dichloro-dihydro-fluorescein diacetate (DCFH-DA) flow cytometry assays, respectively. Western blotting was used to detect mesangial cell expression of PI3K/Akt/mTOR pathway components, including Akt, p-Akt, mTOR, and p-mTOR, and proteins related to cell injury, including TGF-β1 and fibronectin (FN). mRNA expression of TGF-β1 and FN were evaluated using real-time quantitative polymerase chain reaction (PCR). RESULTS Abnormal proliferation was observed in human glomerular mesangial cells at 48 h after treatment with HG, and UA suppressed the HG-induced proliferation of mesangial cells in a dose-dependent manner. UA inhibited ROS generation and oxidative stress in mesangial cells and mitigated mesangial cell injury. Treatment with UA reduced Akt and mTOR phosphorylation levels in mesangial cells exposed to HG (p<0.05 vs. HG) and downregulated protein and mRNA expression of TGF-β1 and FN in these cells (p<0.05 vs. HG). CONCLUSIONS UA attenuated mesangial cell proliferation and ROS generation by inhibiting HG-mediated PI3K/Akt/mTOR pathway activation, thereby ameliorating mesangial cell damage.
研究熊果酸(UA)对高糖(HG)诱导的人肾小球系膜细胞损伤的保护作用,并确定 UA 是否通过抑制 PI3K/Akt/mTOR 通路的激活来抑制细胞增殖和活性氧(ROS)的产生。
用正常葡萄糖(NG 组)、高葡萄糖(HG 组)、甘露醇(甘露醇高渗对照组)或不同浓度(0.5、1.0 和 2.0mmol/L)UA 的高葡萄糖(HG+UA 组)培养人肾小球系膜细胞。通过甲基噻唑基四唑(MTT)和二氯二氢荧光素二乙酸酯(DCFH-DA)流式细胞术分别评估细胞增殖和细胞内 ROS 水平。Western blot 用于检测系膜细胞 PI3K/Akt/mTOR 通路成分的表达,包括 Akt、p-Akt、mTOR 和 p-mTOR,以及与细胞损伤相关的蛋白质,包括 TGF-β1 和纤维连接蛋白(FN)。采用实时定量聚合酶链反应(PCR)评估 TGF-β1 和 FN 的 mRNA 表达。
HG 处理 48 小时后,人肾小球系膜细胞出现异常增殖,UA 呈剂量依赖性抑制 HG 诱导的系膜细胞增殖。UA 抑制了系膜细胞中 ROS 的产生和氧化应激,并减轻了系膜细胞的损伤。UA 处理降低了 HG 暴露的系膜细胞中 Akt 和 mTOR 的磷酸化水平(p<0.05 vs. HG),并下调了这些细胞中 TGF-β1 和 FN 的蛋白和 mRNA 表达(p<0.05 vs. HG)。
UA 通过抑制 HG 介导的 PI3K/Akt/mTOR 通路的激活,减弱了系膜细胞的增殖和 ROS 的产生,从而改善了系膜细胞的损伤。