摘要
目的探讨二苯乙烯苷(TSG)抗动脉粥样硬化(AS)的作用机制,并筛选新的TSG作用靶分子。方法采用高脂饲料喂饲+VitD3复制大鼠动脉粥样硬化模型,随机分为5组:正常对照组(普通饲料组)、模型组(高脂饲料)、TSG低剂量组(30 mg·kg-1·d-1)、TSG中剂量组(60 mg·kg-1·d-1)、TSG高剂量组(120 mg·kg-1·d-1)。造模给药12周后分离大鼠主动脉,提取总蛋白和总RNA。双向凝胶电泳分析差异蛋白质点并初步定量,切割出差异蛋白质点,酶解并进行质谱分析。Western blot检测各组中波形蛋白(差异蛋白质点)表达的变化,RT-PCR检测各组中波形蛋白(差异蛋白质点)转录的变化。结果双向凝胶电泳和质谱分析结果显示,TSG作用后的差异蛋白质为波形蛋白,与正常组相比,AS模型组的波形蛋白呈高表达,TSG作用后波形蛋白表达受到明显的抑制。Western blot和RT-PCR结果表明,TSG对波形蛋白表达的抑制作用可能是在转录水平。结论TSG的抗AS作用很可能与波形蛋白的下调作用有关,波形蛋白有望成为新的TSG作用靶分子。
Aim To investigate the mechanisms of 2, 3,5,4＇-tetrahydroxystilbene-2-O-β-D-glucoside （TSG） for its atherosclerotic effects, and screen novel targets of TSG. Methods Rats were fed with high fat diet and ip vitamin D3 to establish atherosclerosis （AS） model. Rats were divided into the following groups： control group （normal feeds）, model group （high fat feeds）, TSG 30 mg ·kg^-1·d^-1, TSG 60 mg ·kg^-1·d^-1, TSG120mg·kg^-1·d^-1. After l2 weeks , a- ortas of rats were separated. Total protein and RNA were extracted. Differential proteins were analyzed by two-dimensional gel electrophoresis （2-DE） and quan- tified initially. Differential proteins were excised and analyzed by enzymolysis and mass spectroscopy （MS）. Expression and transcription of vimentin were detected by Western blot and RT-PCR analysis, respectively.Results 2-DE and MS analysis indicated that the dif- ferential protein after TSG treatment was vimentin. When compared with the control group, expression of vimentin in AS model group was greatly increased. Af- ter treated with TSG, vimentin expression was declined evidently. Furthermore, according to the results of Western blot and RT-PCR, the inhibitory effect of TSG on vimentin expression might be in the transcription level. Conclusion TSG which could inhibit athero- sclerosis may be associated with the down -regulation of vimentin expression. Vimentin is expected to be- come a novel TSG target.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2013年第9期1260-1264,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81202879)
南通大学人才引进启动费项目(No 122091)
南通市应用研究计划(BK2012087)
南通市科技计划项目(BK2011044)
关键词
二苯乙烯苷
动脉粥样硬化
波形蛋白
主动脉
二维电泳
质谱分析
2,3,5,4＇-tetrahydroxystilbene-2-O-β-D-glucoside
atherosclerosis
vimentin
aorta
two-di-mensional gel electrophoresis
mass spectroscopy
