矢车菊素-3-葡萄糖苷通过降低STAT3活化抑制TNF-α诱导的小鼠血管平滑肌细胞增殖  

Cyanidin-3-glucoside Inhibits TNF-α-Induced Mouse Vascular Smooth Muscle Cells Proliferation through Suppression of STAT3 Activation

作　　者：方仕[1] 罗小琴[2] 吴晓滨[3] 麦海妍[1] 彭俊生[3] 卢味[1] 卓淑雨[1] 叶艳彬[1] 

机构地区：[1]中山大学附属第一医院临床营养科,广东广州510080 [2]中山大学公共卫生学院营养系,广东广州510080 [3]中山大学附属第六医院胃肠外科,广东广州510655

出　　处：《中山大学学报（医学科学版）》2012年第6期716-722,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家"十一.五"科技支撑计划(2008BAD91B00);广东省教育部产学研结合项目(2011B090400558)

摘　　要：【目的】探讨矢车菊素-3-葡萄糖苷(C3G)对TNF-α诱导的血管平滑肌增殖的影响及可能的机制。【方法】C57BL/6J小鼠主动脉平滑肌细胞(VSMC)购于ATCC,体外培养后以C3G对细胞进行预处理后观察TNF-α的促细胞增殖作用;Dihydroethidium(DHE)荧光染色检测VSMC在TNF-α作用下的活性氧(ROS)生成情况;实时定量PCR检测细胞内NADPH氧化酶活化蛋白1(NoxA1)的mRNA水平;蛋白质印迹法检测细胞内NoxA1、信号转导与转录激活因子3(STAT3)、磷酸化STAT3及β-actin的表达水平。统计学方法采用单因素方差分析。【结果】C3G预处理能够抑制TNF-α诱导的VSMC增殖,该作用呈现剂量、时间依赖性。联合应用50μmol/L C3G及100μmol/L apocynin显著减少TNF-α诱导ROS的生成。C3G联合apocynin较C3G或apocynin单独孵育更能显著抑制TNF-α处理下VSMC内NoxA1基因的表达及STAT3蛋白的磷酸化。应用ROS清除剂触媒(catalase 2 000 U/mL)能显著抑制TNF-α诱导的VSMC增殖及STAT3蛋白活化。【结论】C3G对抗TNF-α诱导VSMC增殖的机制很可能是通过削弱NoxA1导致的ROS生成,从而抑制STAT3蛋白的激活。[ Objective ] To investigate the role of Cyanidin-3-glucoside （C3G） in TNF-α-inducd proliferation of vascular smooth muscle cells （VSMC） and the possible mechanisms. [ Methods ] VSMC derived from C57BL/6J mouse aorta were obtained from ATCC and cultured in vitro. The influence of C3G on TNF-α-induced cell proliferation was assessed by cell proliferation assay. The TNF-α- induced production of reactive oxygen species （ROS） in VSMC was evaluated by Dihydroethidium （DHE） staining. The mRNA level of NADPH Oxidase Activator 1 （NoxA1） was measured by real-time PCR. The protein levels of NoxA1, Signal Transducer and Activator of Transcription 3 （STAT3）, Phosphorylated STAT3 and β-actin were semi-quantitatively measured by western blotting. The significance of difference （P 〈 0.05） was determined by one-way ANOVA using SPSS13.0. [Results] C3G pretreatment inhibited VSMC proliferation induced by TNF-α in a dose and time-dependent manner. Combination of 50 μM C3G and 100 μM apocynin significantly reduced TNF-α mediated production of ROS. Combined usage of C3G and apocynin was more effective in suppressing NoxA1 gene expression and STAT3 phosphorylation in VSMC treated by TNF-α than using C3G or apocynin alone. ROS scavenger catalase （2000 U/mL） significantly inhibited TNF-a-induced VSMC proliferation and STAT3 activation. [Conclusions] The mechanism of the anti-proliferating effect of C3G on TNF-α treated VSMC was probably through the repression of NoxA1 induced ROS production and the subsequent inhibition of STAT3 activation.

关 键 词：矢车菊素-3-葡萄糖苷 信号转导与转录激活因子3 活性氧 血管平滑肌细胞 NADPH氧化酶活化蛋白1 

分 类 号：R363[医药卫生—病理学] R543.3[医药卫生—基础医学]