川芎嗪对小鼠巨噬细胞RAW264.7蛋白质组影响的研究  

Study for the Effects of Ligustrazine on Mouse Macrophage RAW264.7 Proteome

作　　者：时小红[1] 卢德赵[1] 宋俊玲[1] 沃兴德[1] 

机构地区：[1]浙江中医药大学生命科学学院,浙江杭州310053

出　　处：《中华中医药学刊》2012年第2期257-259,共3页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(30070932);浙江省中医药管理局重点项目(2006Z002;2006Z2003);浙江省自然科学基金资助项目(Y2100613)

摘　　要：目的:从蛋白质组学水平探讨川芎嗪对巨噬细胞的作用机理。方法:巨噬细胞RAW264.7分成两组:正常对照组、实验组(川芎嗪用药组)。按照实验确定的药物浓度和作用时间作用巨噬细胞后,以差速离心法和超声裂解细胞法提取蛋白质,然后蛋白质双向电泳、质谱仪测定获得肽指纹图谱,ImageMaster 2D 6.0软件进行分析,筛选差异在2倍上下分数的蛋白质点作为有显著意义的蛋白进行质谱鉴定和生物信息学详细分析。结果:实验组与对照组比较,β-肌动蛋白,核纤层蛋白、甘油三磷酸脱氢酶、蛋白酶(前体,macropain)亚基、丙酮酸激酶、钙网织蛋白、肌球蛋白等表达均降低;细胞色素B5、核仁磷蛋白、抗氧化酶Ⅱ等表达升高。结论:川芎嗪很可能通过相关蛋白的调节影响了巨噬细胞的外部形态和运动功能,对其存在的稳定性和内在功能的发挥有重要意义。同时对细胞内部氧化、代谢的参与及凋亡的调节,很可能延长了细胞寿命,从而推测川芎嗪由外及内的调节着巨噬细胞的存在状态。Objective ： It elaborated about biologic1 action mechanism of ligustrazine on macrophage from the angle of proteome. Methods ： Macrophage RAW264.7 were divided into two groups ： the first one was normal control group, the second was experimental group （Ligustrazine treating group）. According to determined concentration of drug and suitable time that was obtained by experimentations, proteins were extracted by differential centrifugation and ultrasound spallation method. Then two - dimensional electrophoresis and mass spectrum were used to study the protein differences of normal control group and the experimental group for gaining peptid finger printing. Then the gels were respectively imaged by UMAX Image Scanner and spot - features of proteome difference were analyzed by ImageMaster 2D 6. 0 software. MALD I - TOFMS was used to identify the different protein which abundance changed more than 2 folder up or down between normal control group and experimental group. Results ： In the experimental group, the expression of β - actin protein, Lamin protein, three glycerol phosphate dehydrogenase protein, protease （ precursors, macropain） subunit protein, PK protein, calreticulin and myosin protein decreased ; cytochrome B5, nucleolar phosphorus protein and antioxidant enzymes II protein increased. Conclution ： The macrophages＇ external morphology and motor function were regulated by ligu strazine, because some proteins had changed which was very significant in aspects of cell＇s stability and function. As the participation of intracellular oxidation, metabolism and apoptosis, which was likely to lead to prolong cell life, so, from the above all it suggested that ligustrazine regulated macrophages from outside to inside.

关 键 词：川芎嗪 巨噬细胞 蛋白质组 

分 类 号：R285.5[医药卫生—中药学]