茶多酚对鼠胎儿成纤维细胞凋亡的影响  

Effects of tea polyphenols on the apoptosis of mouse embryonic fibrolast

作　　者：王洋[1] 盛宇[1] 李钟淑[1] 方南洙[1] 金一[1] 

机构地区：[1]延边大学农学院,吉林延吉133002

出　　处：《中国农业大学学报》2015年第2期149-155,共7页Journal of China Agricultural University

基　　金：国家自然科学基金资助项目(31360546);吉林省科技厅重点项目(20100228)

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150μg/mL的TP,分别培养24、48、72和96h,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72h,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40μg/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100μg/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150μg/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100μg/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。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 150μg/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-3was 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 100μg/mL,the growth of MEF（P〈0.05）was inhibited and the cell morphology had no different changes.At the concentration of 40μg/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 150μg/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 Baxgene,Bcl-2 gene and Caspase-3 gene（P〉0.05）.When the TP concentration was higher than 100μg/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.

关 键 词：茶多酚 鼠胎儿成纤维细胞 细胞凋亡 反转录 

分 类 号：Q813[生物学—生物工程] S865.13[农业科学—野生动物驯养]