Resveratrol protects vascular smooth muscle cells against high glucose-induced oxidative stress and cell proliferation in vitro.
Resveratrol protects vascular smooth muscle cells against high glucose-induced oxidative stress and cell proliferation in vitro.
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DOI：
10.12659/msmbr.890858
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发表时间：
2014-06-27
影响因子：
2.8
通讯作者：
Xu Y
中科院分区：
文献类型：
--
作者：
Guo R;Li W;Liu B;Li S;Zhang B;Xu Y
白藜芦醇在发病的不同阶段对许多退行性疾病表现出有益的作用。本研究探讨了潜在的机制和白藜芦醇对高糖（HG）诱导的氧化应激（30 mM D-葡萄糖，30分钟）和细胞增殖（30 mM D-葡萄糖，24小时）在血管平滑肌细胞（VSMCs）的影响。用2′，7 ′-二氯荧光素二乙酸酯（DCFH-DA）检测细胞内活性氧（ROS）的产生。测定总抗氧化能力（TAC）、丙二醛（MDA）、谷胱甘肽（GSH）和超氧化物歧化酶（SOD）以评价氧化应激。CCK-8法和碘化丙啶细胞周期分析法检测VSMC增殖。通过免疫印迹分析评估NAD（P）H氧化酶、增殖蛋白和细胞信号传导的表达。1-100 μM白藜芦醇可降低HG诱导的VSMCs活性氧的产生（P<0.05）。白藜芦醇还废除了汞诱导的氧化酶亚基p47 phox的磷酸化和减少汞诱导的细胞周期蛋白D1，细胞周期蛋白E，和PCNA的表达浓度依赖性的方式。此外，白藜芦醇（10 μM）减弱HG诱导的Akt、p38丝裂原活化蛋白激酶（MAPK）、ERK 1/2和JNK 1/2磷酸化，而不影响总水平。HG刺激增强下游IκB-α磷酸化和NF-κB活性，白藜芦醇抑制这些作用。白藜芦醇通过抑制ROS生成、NADPH氧化酶、Akt磷酸化、p38 MAPK/JNK/ERK磷酸化以及IκB-α和NF-κB活性，抑制HG诱导的氧化应激和VSMC增殖。
Resveratrol exhibits beneficial effects against numerous degenerative diseases at different stages of pathogenesis. This study investigated potential mechanisms and resveratrol effects on high glucose (HG)-induced oxidative stress (30 mM d-glucose, 30 min) and cell proliferation (30 mM d-glucose, 24 h) in vascular smooth muscle cells (VSMCs). Intracellular reactive oxygen species (ROS) generation was detected by 2′,7′-dichlorofluorescein diacetate (DCFH-DA). Total antioxidant capacity (TAC), malonyldialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) were measured to evaluate oxidative stress. VSMC proliferation was measured by CCK-8 assays and through propidium iodide-based cell cycle analysis. Expression of NAD(P)H oxidase, proliferation proteins, and cell signalling were assessed by immunoblot analysis. Co-treatment of primary cultures of VSMCs with 1–100 μM resveratrol decreased HG-induced ROS overproduction (P<0.05). Resveratrol also abolished HG-induced phosphorylation of oxidase subunit p47 phox and reduced HG-induced cyclin D1, cyclin E, and PCNA expression in a concentration-dependent manner. Furthermore, resveratrol (10 μM) attenuated HG-induced phosphorylation of Akt, p38 mitogen-activated protein kinase (MAPK), ERK 1/2, and JNK1/2 without affecting total levels. HG stimulation enhanced downstream IκB-α phosphorylation and NF-κB activity, and resveratrol repressed these effects. Resveratrol inhibits HG-induced oxidative stress and VSMC proliferation by suppressing ROS generation, NADPH oxidase, Akt phosphorylation, p38 MAPK/JNK/ERK phosphorylation, and IκB-α and NF-κB activities.