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作 者:王继生[1] 邱宗荫[2] 夏永鹏[2] 李惠芝[2]
机构地区:[1]绵阳市第三人民医院,四川绵阳621000 [2]重庆医科大学药学院,重庆400016
出 处:《中国药理学通报》2009年第2期201-204,共4页Chinese Pharmacological Bulletin
基 金:重庆市重大科技专项资助课题[No渝科发计字(2004)27号]
摘 要:目的采用蛋白质组学技术研究茅莓总皂苷对脑缺血/再灌注大鼠脑组织差异蛋白的影响,进一步揭示茅莓总皂苷的脑保护作用及抗损伤修复机制。方法建立脑缺血/再灌注大鼠动物模型,分别提取蛋白质后进行2-DE电泳分离,应用相关软件比较分析,找出差异蛋白,酶切消化后进行肽指纹图谱分析,通过数据检索,初步鉴定差异蛋白质。结果与模型组比较,茅莓总皂苷组找到29个差异点,其中18个蛋白点表达上调,11个蛋白点表达下调,鉴定了其中7个蛋白。结论茅莓总皂苷对脑保护作用及抗损伤修复机制的机制是多个蛋白质共同参与的结果。Aim To screen alteration in protein level in brain tissue between middle cererbral artery occlusion (MCAO) and the total saponins of Rubus Parviflolius L(TSRP) treated rats and to learn the mechanism of nerve-protecting effects of TSRP after focal cerebral ischemia-reperfusion. Methods The MCAO 2 h reperfusion of 24 h model was established in rats,2-DE was applied to preparein step 1 for the separation of protein, and imaging analysis of gels was done with Image-Master 2D software. Different protein spots expressing between cerebral ischemic reperfusion (CIR) model and TSRP group were cut from the gels for the MS analysis, Different protein spots expressing between cerebral ischemie tissue administered with or without TSRP were also cut from the gels for the MS analysis.MALDI-TOF-MS and database search: protein spots from step 2 were subject to in - gel digestion and MAL- DI-TOF-MS analysis, which resulted in the peptide mass fingerprint spectra (PMF). PMF were search against swiss-prot database using search program of Ms- Fit. Results By comparative proteome analysis, 29 expression-altered spots were excised for MS and informatics analysis, comparing to the CIR model, 18 proteins up-regulated in the TSRP group, 11 proteins down-regulated, 7 spots of them were identified and characterized. Conclusion The protective effect of TSRP on cerebral ischemia-reperfusion is the result of the participation of a number of proteins.
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