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目的:研究夏枯草提取物作用于Jurkat细胞后蛋白质组的变化。方法:体外培养Jurkat细胞,用MTT法观察不同浓度夏枯草提取物对细胞的增殖抑制作用。加入20μg/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,实现对蛋白点的定性鉴定。结果:夏枯草提取物可显著抑制Jurkat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11个蛋白质,包括glyceraldehyde-3-phosphate dehydrogenase、coagulation factor VII、Heterogeneous nuclear ribonucleoprotein L、heat shock 70 kDa protein 8 isoform 2、immunoglobulin heavy chain variable region、heterogeneous nuclear ribonucleoprotein A2/B1、heterogeneous nuclear ribonucleoprotein L(为heterogeneous nuclear ribonucleoprotein A2/B1不同亚型)、zinc finger protein 43、chaperonin containing TCP1、subunit 6A (zeta 1)、isoform CRA-b。结论:夏枯草提取物可显著抑制Jurkat细胞的生长,并引起Jurkat细胞蛋白质组的改变,这可能是夏枯草提取物抗肿瘤作用的机制之一。
OBJECTIVE: To study the changes in the proteome of Jurkat cells treated with Prunella vulgaris extract. METHODS: Jurkat cells were cultured in vitro and MTT assay was used to observe the inhibitory effect of different concentrations of Prunella vulgaris extract on cell proliferation. 20 μg/mL Prunella vulgaris extract was added to the cells for 48 h. The total protein was extracted and analyzed by two-dimensional electrophoresis. The gel gel was stained with silver and analyzed by ImageMaster 2D Platium 5.0 software to find differentially expressed proteins. The difference points were cut, and the MALDI-TOF-MS analysis and database search were performed after in-gel digestion to achieve the qualitative identification of protein spots. Results: Prunella vulgaris extract can significantly inhibit the growth of Jurkat cells, and has a certain dose-effect relationship. After two-dimensional electrophoresis and mass spectrometry, 11 proteins were successfully identified, including glyceraldehyde-3-phosphate dehydrogenase, coagulation factor VII, Heterogeneous nuclear ribonucleoprotein L, heat shock 70 kDa protein 8 isoform 2, immunoglobulin heavy chain variable region, and heterogeneous nuclear ribonucleoprotein A2. /B1. Heterogenous nuclear ribonucleoprotein L (different isoforms of heterogeneous nuclear ribonucleoprotein A2/B1), zinc finger protein 43, chaperonin containing TCP1, subunit 6A (zeta 1), isoform CRA-b. Conclusion: Prunella vulgaris extract can significantly inhibit the growth of Jurkat cells and cause changes in the proteome of Jurkat cells, which may be one of the mechanisms of the anti-tumor effect of Prunella vulgaris extract.