调控胰岛素致自发性高血压大鼠VSMC增殖的蛋白  

Identification of proteins involved in insulin stimulation in vascular smooth muscle cells of SHR rats

作　　者：王旭开[1] 王燕[2] 杨成明[1] 付春江[1] 张晔[1] 何作云[3] 

机构地区：[1]第三军医大学大坪医院心血管内科,重庆400042 [2]第三军医大学分子遗传教研室,重庆400042 [3]第三军医大学新桥医院心血管内科,重庆400042

出　　处：《中国病理生理杂志》2006年第6期1061-1065,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30170384)

摘　　要：目的:探讨胰岛素对自发性高血压大鼠血管平滑肌细胞(VSMC)增殖调控中参与调控的蛋白质。方法:分离、培养SHR、WKY大鼠的VSMC,各分对照组、胰岛素组。双向电泳技术分离样品,对差异的蛋白质点进行肽质量指纹测定。差异的蛋白质用免疫印迹验证,同时观察胰岛素干预前后VSMC的增殖率和迁移率。结果:SHR大鼠胰岛素组的VSMC[3H]-TdR掺入值(counts/min)14·40±0·85高于WKY大鼠胰岛素组的VSMC[3H]-TdR掺入值(counts/min)9·21±0·93。SHR大鼠胰岛素组VSMC的迁移率是WKY大鼠胰岛素组的1·8倍(P<0·05)。双向电泳分析SHR大鼠胰岛素组与对照组相比有312个点相匹配。19个蛋白质点产生较好的质谱图,其中14个点与已知蛋白质相匹配。免疫印迹证实SHR VSMC中基质蛋白(matrix gla)和骨桥蛋白(OPN)在胰岛素干预后升高,而肌动蛋白(α-SM)降低。WKY大鼠与SHR大鼠不同的是胰岛素干预后VSMC中matrix gla、OPN和肌动蛋白变化不显著。结论:胰岛素刺激后差异蛋白可能在不同遗传背景的大鼠VSMC异常增殖中具有不同的表现。AIM： To identify proteins involved in insulin stimulation and the molecular mechanism of proliferation and migration in vascular smooth muscle cells （VSMCs）. METHODS： A series of methods, including 2 - D electrophoresis, PDQuest software analysis of 2 - DE gels, peptide mass fingerprinting based on matrix - assisted laser desorption/ionization time of flight mass spectrometry （MALDI - TOF- MS） and SWISS- PROT database searching, were used to separate and identify the differentially expressed proteins. The difference of some proteins was proved by Western blotting. Proliferation and migration of VSMCs treated with insulin were also observed. RESULTS： DNA synthesis were increased in VSMCs. [^3H] - thymidine incorporation in VSMCs from SHR （14.40±0.85） was higher than that in VSMCs from WKY （9.21±0.93, P 〈 0.05）. Migration rate of VSMCs from SHR was about 1.8 times higher than that in VSMCs from WKY （P 〈 0.05）. Image analysis revealed that averages of protein spots detected were 502 ± 32 and 612 ± 39 in control VSMCs and in cells treated with insulin, respectively. Result of western blotting confinned that α- SM protein was downregulated and matrix Gla, OPN proteins were upregulated by insulin stimulation. CONCLLISION： The results suggest that the differential proteomic analysis may be useful to study related proteins involved in insulinregulated proliferation in VSMCs.

关 键 词：血管平滑肌细胞 胰岛素 电泳 表型 

分 类 号：R363[医药卫生—病理学]