Vaspin attenuates high glucose-induced vascular smooth muscle cells proliferation and chemokinesis by inhibiting the MAPK, PI3K/Akt, and NF-κB signaling pathways
Vaspin attenuates high glucose-induced vascular smooth muscle cells proliferation and chemokinesis by inhibiting the MAPK, PI3K/Akt, and NF-κB signaling pathways
复制标题
DOI：
10.1016/j.atherosclerosis.2013.02.013
复制
发表时间：
2013-05-01
期刊：
影响因子：
5.3
通讯作者：
Xu, Yawei
中科院分区：
文献类型：
--
作者：
Li, Hailing;Peng, Wenhui;Xu, Yawei
背景：Vaspin具有胰岛素增敏作用，以及对代谢性疾病的额外有益作用。然而，关于血管素对糖尿病介导的血管并发症的直接作用，目前所知甚少。本研究旨在探讨vaspin对高血糖诱导的血管平滑肌细胞（VSMCs）增殖、趋化运动和细胞信号传导的影响及其机制。方法：采用5-乙基-2'-脱氧尿苷细胞增殖法检测大鼠VSMCs增殖，采用划痕法监测趋化动力学，采用H(2)DCFDA法和sod抑制铁细胞色素c还原法检测活性氧（ROS）生成。荧光素酶活性使用双荧光素酶报告检测系统进行检测。通过免疫印迹法评估细胞信号。结果：Vaspin显著抑制高糖（HG）诱导的VSMCs增殖、趋化、ROS生成和NADPH氧化酶活性。与HG相比，100 ng/ml vaspin显著降低VSMCs增殖40 +/- 8%。Vaspin在100 ng/ml和300 ng/ml浓度下也分别使ROS生成降低16 +/- 8%和30 +/- 8%（均P < 0.01）。Vaspin在不影响氧化酶亚基p47phox、Akt、p38和JNK1/2总水平的情况下，显著消除hg诱导的氧化酶亚基p47phox、Akt、p38和JNK1/2的磷酸化，并减弱hg诱导的胰岛素受体及其下游IRS-1和IRS-2的磷酸化。对于下游靶点，HG刺激后NF-kappa B活性和I kappa B α磷酸化均显著增强，而这些作用被vaspin抑制。Vaspin还能显著抑制hg诱导的PCNA和cyclin D1的表达。结论：Vaspin通过抑制ROS激活和MAPK、PI3K/Akt和NF-kappa B信号传导抑制hg诱导的VSMCs增殖和趋化。2013爱思唯尔爱尔兰有限公司版权所有。
Background: Vaspin has insulin-sensitizing effects, as well as additional beneficial effects on metabolic diseases. However, little is known about the direct effects of vaspin on vascular complications mediated by diabetes. The objective of this study is to determine the efficacy and mechanism of vaspin on hyperglycemia-induced vascular smooth muscle cells (VSMCs) proliferation, chemokinesis and cell signaling.Methods: Rat VSMCs proliferation was determined with 5-ethynyl-2'-deoxyuridine cell proliferation assays, chemokinesis was monitored with scratch assays, and reactive oxygen species (ROS) production was assessed using H(2)DCFDA and SOD-inhibited reduction of ferricytochrome c assay. Luciferase activity is assayed using a Dual Luciferase Reporter Assay System. Cell signaling is assessed by immunblotting.Results: Vaspin significantly inhibited VSMCs proliferation and chemokinesis, as well as ROS generation and NADPH oxidase activity, induced by high glucose (HG) treatment. Compared with HG, vaspin significantly decreased VSMCs proliferation by 40 +/- 8% at 100 ng/ml. Vaspin also decreased ROS production by 16 +/- 8% at 100 ng/ml and 30 +/- 8% at 300 ng/ml (all P < 0.01). Vaspin significantly abolished HG-induced phosphorylation of oxidase subunits p47phox, Akt, p38, and JNK1/2 without affecting their total levels, and attenuated HG-induced phosphorylation of insulin receptor and its downstream IRS-1 and IRS-2. For downstream targets, NF-kappa B activity and I kappa B alpha phosphorylation were both enhanced significantly after HG stimulation, and these effects were inhibited by vaspin. Vaspin also significantly abolished HG-induced PCNA and cyclin D1 expression.Conclusions: Vaspin inhibits HG-induced VSMCs proliferation and chemokinesis by preventing ROS activation and MAPK, PI3K/Akt, and NF-kappa B signaling. (C) 2013 Elsevier Ireland Ltd. All rights reserved.