木犀草素对小鼠T淋巴细胞体外增殖及凋亡的作用

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(P0.01),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(P0.01),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(P0.01),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。

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

Objective:To investigate the effect of luteolin on the proliferation and apoptosis of T cells in vitro. Method:Blank group,ConA 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 ConA 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.