局灶性脑缺血大鼠海马蛋白质组差异表达图谱的构建及针刺影响

Effect of acupuncture and construction of differential expression map of hippocamp proteome in rats with focal cerebral ischemia

[目的]构建局灶性脑缺血早期大鼠海马蛋白质组差异表达图谱,寻找与脑缺血相关蛋白质,探讨脑缺血的病理机制及针刺影响。[方法]Wistar大鼠随机分为正常组、假手术6 h、模型6 h、醒脑开窍针刺6 h(简称醒针6 h)和传统对照6 h组(简称对照6 h),采用改进的线栓法制备动物模型。在规定时间快速断头取脑,剥离缺血侧海马,提取脑组织总蛋白进行双向电泳,以Image Master 2DV3.01 Elite软件进行图像分析。选取差异蛋白质点进行胶内酶解,以(MALDI-TOF-MS)测肽质量指纹图,检索Swiss-Port数据库,对蛋白质进行鉴定。[结果]正常组与假手术组比较无差异蛋白表达;模型组与正常组比较鉴定出差异蛋白质点29个,其中在模型6 h组新出现1个,在模型6 h表达上调的11个,表达下调的17个;醒脑开窍针刺组与模型组比较鉴定出差异蛋白质点21个,其中在醒脑开窍针刺组上调有16个,下调的有5个;对照组与模型组比较鉴定出差异蛋白质点20个,下调的有8个,上调的有12个。[结论]以双向电泳联合质谱技术,初步发现与脑缺血相关的部分蛋白质,有助于深入研究脑缺血性损伤病理机制,并发现针刺治疗脑缺血的部分靶蛋白质,及"醒脑开窍"针刺法与传统对照针刺治疗脑缺血性损伤作用机制的异同。

[Objective] To construct differential expression map of hippocamp proteome in rats with focal cerebral ischemia to search the related protein of cerebral ischemia and explore the pathogenesis mechanism of cerebral ischemia and the effect of acupuncture. [Methods] Wistar rats were randomly divided into normal group, sham operation group, model group, Xingnao Kaiqiao （restoring consciousness and waking up the patient from unconsciousness） acupuncture group and traditional acupuncture control group. Focal cerebral ischemia was induced by line-bolted method in rats. Image analysis by Image Master 2D V3.01 Elite software was carried out and the finger print map of peptide quality by MALDI-TOF-MS method was measured. The Swiss Port database was retrieved and protein appraisement was carried out. [Results] There wasn＇t differential protein expression in normal group and sham operation group. Compared with the normal group twenty-nine differential protein particles was identified in model group. The differential protein particles included one particle emerged in 6 h group, eleven particles with upregulating expression and seventeen particles with downregulating. Compared with model group twenty-one differential protein particles in Xingzhen group were identified including sixteen particles with upregulating expressing and five particles with downregulating. Compared with model group twenty differential protein particles in acupuncture control group were identified including twelve particles with upregulating expression and eight particles with downregulating. [Conclusions] The partial related protein with cerebral ischemia was discovered by two-dimensional electrophoresis associated with mass spectra technique. The study result is helpful for lucubrating the pathogenesis mechanism of cerebral ischemia injury and discovering the effect mechanism of partial target protein in treating cerebral ischemia.