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目的:分析夏枯草提取物作用于Raji细胞后蛋白质组的变化。方法:体外培养Raji细胞,用MTT观察不同浓度夏枯草提取物对细胞的增殖抑制作用。加入18μg/mL夏枯草提取物作用于细胞48 h,提取总蛋白,进行双向电泳测定,凝胶银染显色,用ImageMaster 2D Platium 5.0软件对获得的蛋白图谱加以分析,寻找差异表达的蛋白质。切取差异点,胶内酶切后进行MALDI-TOF-MS分析和数据库搜索,实现对蛋白点的鉴定。结果:夏枯草提取物可显著抑制Raji细胞的生长,且具有一定的量效关系。经双向电泳和质谱后,成功鉴定了27个(已知蛋白22个,未知蛋白5个)蛋白质,包括macrophin 1 isoform 2、mitochondrial heatshock 60×103protein 1 variant 1、similar toPIK4CA variant protein、glyceraldehyde-3-phos-phate dehydrogenase、chaperonin containing TCP1、subunit 2(beta)、isoform CRA_a、methylcrotonoyl-Coenzyme A carboxylase 2(beta)、chaperonin con-taining TCP1、subunit 6A(zeta 1)和isoformCRA_b等。结论:夏枯草提取物可显著抑制Raji细胞的生长,并引起Raji细胞蛋白质组的改变,可能与夏枯草提取物的抗肿瘤作用有关。
Objective: To analyze the proteome changes of Prunella vulgaris extract on Raji cells. Methods: Raji cells were cultured in vitro. MTT assay was used to observe the inhibitory effects of different concentrations of Prunella vulgaris extract on cell proliferation. After adding 18μg / mL Prunella vulgaris extract to the cells for 48 h, the total protein was extracted and analyzed by two-dimensional gel electrophoresis and gel silver staining. The protein profiles obtained were analyzed by ImageMaster 2D Platium 5.0 software to find differentially expressed proteins. Cut the differences, in-gel digested MALDI-TOF-MS analysis and database search, the identification of protein spots. Results: Prunella vulgaris extract can significantly inhibit the growth of Raji cells, and has a certain dose-effect relationship. After two-dimensional electrophoresis and mass spectrometry, 27 proteins (22 known proteins and 5 unknown proteins) were successfully identified, including macrophin 1 isoform 2, mitochondrial heatshock 60 × 103 protein 1 variant 1, similar to PIK4CA variant protein, glyceraldehyde- phos-phate dehydrogenase, chaperonin containing TCP1, subunit 2 (beta), isoform CRA_a, methylcrotonoyl-Coenzyme A carboxylase 2 (beta), chaperonin con taining TCP1, subunit 6A (zeta 1) and isoformCRA_b. Conclusion: Prunella vulgaris extract can significantly inhibit the growth of Raji cells and cause proteomic changes in Raji cells, which may be related to the antitumor effect of Prunella vulgaris extract.