松油烯-4-醇干预KLF4信号改善高糖诱导VSMCs氧化应激损伤的实验研究  

Effect of terpinen-4-ol ameliorating oxidative stress in VSMCs by down-regulation of KLF4

作　　者：何丽 黄梅 杨彩琴 陈娜 付凌云[2] 蒋朝晖 王惠林 沈祥春[2] 徐旖旎[2] 张彦燕 HE Li;HUANG Mei;YANG Cai-qin;CHEN Na;FU Ling-yun;JIANG Zhao-hui;WANG Hui-lin;SHEN Xiang-chun;XU Yi-ni;ZHANG Yan-yan(Dept of Clinical Pharmacy,School of Pharmacy,Guizhou Medical University,Guiyang 550025,China;Key Laboratory of Optimal Utilization of Natural Medicine Resources,Guizhou Medical University,Guiyang 550025,China;The First People’s Hospital of Guiyang,Guiyang 550002,China;The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China)

机构地区：[1]贵州医科大学药学院临床药学教研室,贵州贵阳550025 [2]贵州医科大学天然药物资源优效利用重点实验室,贵州贵阳550025 [3]贵州省第一人民医院,贵州贵阳550002 [4]贵州中医药大学第一附属医院,贵州贵阳550001

出　　处：《中国药理学通报》2020年第6期839-844,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金(No 81560811);国家自然科学基金喀斯特中心项目(No U1812403-4-4);贵州医科大学药学国际科技合作基地(黔科合平台人才[2017]5802);天然药物药理与成药性评价重点实验室(筑科合同[2017]5-21号);贵州省科学技术基金(黔科合基础2016[1128]);贵州省研究生科研基金(黔教合YJSCXJH(2019)074)。

摘　　要：目的探讨松油烯-4-醇(terpinen-4-ol,T4O)对高糖诱导的血管平滑肌细胞(vascular smooth muscle cells,VSMCs)氧化应激损伤的保护作用及可能机制。方法采用高糖(HG,24.5 mmol·L-1)刺激VSMCs细胞,不同剂量T4O预处理后,应用噻唑蓝(MTT)法检测细胞活力,荧光探针2′、7′-二氯荧光素二乙酸酯(DCFH-DA)检测活性氧(ROS)水平,蛋白免疫印迹法检测KLF4、Nrf2和HO-1的表达水平。进一步采用siRNA技术实现KLF4的敲低表达,检测细胞活力、ROS水平,及其对Nrf2、HO-1蛋白表达的影响。结果松油烯-4-醇能够抑制高糖诱导的VSMCs异常增殖,降低ROS水平,下调高糖诱导VSMCs中KLF4的表达,并上调抗氧化信号Nrf2、HO-1的表达。进一步研究表明,siKLF4转染组明显下调KLF4表达,抑制VSMCs增殖,降低细胞内ROS水平,并上调Nrf2、HO-1的表达水平。结论松油烯-4-醇对高糖诱导的VSMCs氧化应激损伤具有保护作用,其机制与其抑制核转录因子KLF4的表达,上调Nrf2/HO-1抗氧化信号轴有关。Aim To investigate the protective effect of terpinen-4-ol(T4O)on oxidative stress injury induced by high glucose in vascular smooth muscle cell(VSMCs),and to explore its possible mechanism.Methods The model of VSMCS oxidative stress injury was established by high glucose(HG,24.5 mmol·L-1).Terpinen-4-ol pretreatment and MTT assay were used to detect cell proliferation,ROS levels were detected by DCFH-DA,and Western blot was used to detect KLF4,Nrf2 and HO-1 expression levels;siRNA technology interfered with KLF4 expression was used to analyze cell viability and ROS levels and its effect on Nrf2,HO-1 and KLF4 protein expression was further assessed.Results T4O could inhibit the abnormal proliferation of VSMCs induced by high glucose.In addition,it could reduce the ROS level.Moreover,it could down-regulate the expression of KLF4 in model cells and up-regulate the expression of Nrf2 and HO-1.In high glucose environment,siRNA interfered with KLF4 expression could reduce VSMCs proliferation and ROS levels,significantly down-regulate KLF4 expression and up-regulate Nrf2,HO-1 expression.Conclusions T4O can ameliorate VSMCs oxidative stress injury status induced by high glucose,and its mechanism may be related to the inhibition of the expression of nuclear transcription factor KLF4 and up-regulating the Nrf2/HO-1 antioxidant signaling axis.

关 键 词：松油烯-4-醇 血管平滑肌细胞 氧化应激 活性氧 Nrf2/HO-1 KLF4 

分 类 号：R-332[医药卫生] R284.1