摘要: |
为探究茶多酚(Tea polyphenols,TP)对体外培养鼠胎儿成纤维细胞(Mouse embryonic fibrolast,MEF)凋亡的影响。在MEF细胞培养液中添加质量浓度为0、10、40、70、100和150 μg/mL的TP,分别培养24、48、72和96 h,使用MTT法(3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐,噻唑蓝,MTT法)检测细胞活力;细胞培养72 h,以RT-PCR检测B-细胞淋巴瘤/白血病-2原癌基因(B cell lymphoma leukemia- 2,Bcl-2)、和Bcl相关的蛋白X(Bcl-as-sociated X protein,Bax)、半胱天冬酶-3(Caspase-3)的相对表达量。MTT结果表明:TP质量浓度为40 μg/mL 时细胞密度略高于其他组,但是没有显著差异(P>0.05);质量浓度为100 μg/mL时,对MEF的生长产生抑制作用(P<0.05),细胞形态没有发生明显变化;当质量浓度为150 μg/mL时对细胞生长抑制作用明显(P<0.05),从形态观察细胞出现调亡;通过反转录结果表明,对Bax基因、Bcl-2基因和Caspase-3基因的分子转录水平没有影响(P>0.05),表明质量浓度高于100 μg/mL TP对MEF生长产生抑制作用,但对Bax基因、Bcl-2基因和Caspase-3转录分子机制无影响。 |
关键词: 茶多酚 鼠胎儿成纤维细胞 细胞凋亡 反转录 |
DOI:10.11841/j.issn.1007-4333.2015.02.019 |
分类号: |
基金项目:国家自然科学基金资助项目(31360546); 吉林省科技厅重点项目(20100228) |
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Effects of tea polyphenols on the apoptosis of mouse embryonic fibrolast |
WANG Yang, SHENG Yu, LI Zhong-shu, FANG Nan-zhu, JIN Yi
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Agricultural College of Yanbian University, Yanji 13002, China
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Abstract: |
In order to explore the effects of tea polyphenols (TP)on the apoptosis of mice fetal fibroblast cells (MEF) culture in vitro, MEF cells were cultured, adding 0, 10, 40, 70, 100 and 150 μg/mL concentrations of TP in the MEF culture fluid and using MTT method, the cell viability was assayed.After 72 hours of the cultured cell, using RT-PCR, the transcription of gene Bcl-2 (B cell lymphoma leukemia-2), Bax (Bcl-as-sociated X protein) and Caspase-3 was tested.Using the beta-actin gene as internal reference, the relative expression amount was tested.The rulsults of MTT mothed(3-(5-Dimethylthiazol-2-yl)-25-diphenyltetrazoliumbromide), showed that when the concentration of TP was 100 μg/mL, the growth of MEF(P<0.05) was inhibited and the cell morphology had no different changes.At the concentration of 40 μg/mL, the result was a bit higher than in other groups, but there were no significant differences(P>0.05).When the TP concentration was 150 μg/mL, the inhibition founction was significant(P<0.05), and the cell morphological had changes.The RT-PCR results showed that TP had no effect on the molecular transcription level of Bax gene, Bcl-2 gene and Caspase-3 gene(P>0.05).When the TP concentration was higher than 100 μg/mL, the growth of MEF was inhibited, and there was nothing to do with gene Bax, gene Bcl-2 and Caspase-3 transcription molecular mechanism. |
Key words: TP MEF apoptosis RT-PCR |