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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: To explore the mechanism of ligustrazine on macrophages from proteomics level. Methods: RAW264.7 macrophages were divided into two groups: normal control group and experimental group (ligustrazine group). The macrophages were treated with the drug concentration and action time determined experimentally, then the proteins were extracted by differential centrifugation and sonication. Then the protein fingerprints were obtained by two-dimensional electrophoresis and mass spectrometry. ImageMaster 2D 6.0 software was used to analyze the differences The protein spots at up and down 2 times were used as mass spectrometry identification and bioinformatics analysis as significant proteins. Results: Compared with the control group, β-actin, lamin, glycerol triphosphate dehydrogenase, protease (macropain) subunit, pyruvate kinase, calcium reticulin, myosin and so on The expression of cytochrome B5, nucleophosmin and proteinase Ⅱ were increased. CONCLUSION: Tetramethylpyrazine may affect the external morphology and motor function of macrophages through the regulation of related proteins, which is of great significance to its stability and intrinsic function. Simultaneously, the internal oxidation and metabolism of cells and the regulation of apoptosis are likely to prolong the cell life. Therefore, it is supposed that ligustrazine can regulate the existence of macrophages from the outside and inside.