木犀草素对小鼠T淋巴细胞体外增殖及凋亡的作用

Effect of Luteolin on Proliferation and Apoptosis of Mouse T Cells in Vitro

目的：研究木犀草素对小鼠T淋巴细胞体外增殖及凋亡作用的影响。方法：设置刀豆蛋白A（Con A）组和空白组,设置3个木犀草素药物组,终质量浓度分别为2.5,5,10 mg·L-1,细胞毒性活性检测-8（CCK-8）法检测各组细胞增殖情况,末端脱氧核苷酸转移酶介导的d UTP缺口末端标记测定法（TUNEL）和Annexin V-FITC/PI双染法检测各组细胞凋亡情况,蛋白免疫印迹法（Western blot）检测各组细胞凋亡通路蛋白B淋巴细胞瘤-2（B-cell lymphoma-2,Bcl-2）,Bcl-2相关X蛋白（Bcl-2-associated X protein,Bax）表达水平。结果：CCK-8实验结果显示3个木犀草素药物组细胞数明显低于Con A组和空白组（P〈0.01）,差异有统计学意义。TUNEL实验结果显示,Con A组的凋亡细胞数量显著低于3个木犀草素药物组（P〈0.01）,差异有统计学意义。流式细胞仪检测结果显示3个木犀草素药物组的细胞凋亡率显著高于Con A组和空白组（P〈0.01）,细胞凋亡率随着药物浓度增加而增高,呈明显的量-效关系。Western blot检测结果显示,2.5,5 mg·L-1木犀草素药物组中Bcl-2表达量高于对照组,Bax表达量低于对照组;10 mg·L-1药物组中Bcl-2表达量低于对照组,Bax表达量高于对照组。结论：2.5,5,10 mg·L-1木犀草素能够抑制T淋巴细胞增殖,能够诱导T淋巴细胞凋亡,10 mg·L-1木犀草素可能主要通过下调Bcl-2,上调Bax影响线粒体凋亡通路诱导细胞凋亡,2.5,5 mg·L-1木犀草素可能通过其他通路诱导细胞凋亡。

Objective： To investigate the effect of luteolin on the proliferation and apoptosis of T cells in vitro. Method： Blank group,Con A control group and 3 luteolin groups（2. 5,5,10 mg·L-1） were set up. The effect of luteolin on the proliferation of T lymphocyte in the 5 groups was detected by cell counting kit-8（CCK-8）.TUNEL and Flow Cytometry were used to detect apoptosis condition in each group. Western blot method was used to detect the expression of B-cell lymphoma-2（Bcl-2） and Bcl-2 associated X protein（Bax） in each group.Result： Number of T cell in 3 different doses of luteolin were decreased significantly compared with 2 control groups（P〈0. 01）,with statistically significant differences. TUNEL and flow cytometry indicated that,compared with Con A groups,apoptotic percentages of the 3 luteolin groups were increased under fluorescence microscope（P〈0. 01）,with statistically significant differences. Among apoptosis-related proteins,Bax was up-regulated,but Bcl-2 was down-regulated in 10 mg·L-1 group according to Western blot. Conclusion： The 2. 5,5,10 mg·L-1 of luteolin can inhibit the proliferation of T cell and induce its apoptosis. 10 mg·L-1 of luteolin may mainly induce the apoptosis by down-regulating expression of Bcl-2 and up-regulating expression of Bax in mitochondria apoptosis pathway,and 2. 5,5 mg·L-1 of luteolin may induce the apoptosis through other pathways.