雪菊不同极性萃取物对小鼠脾脏CD4^+ T细胞体外增殖的影响

Effects of different polar extracts of Coreopsis tinctoria on proliferation of spleen-derived CD4^+ T cells of mouse in vitro

目的:观察不同极性雪菊萃取物对小鼠CD4+T细胞体外增殖的影响。方法:用95%乙醇和不同极性溶剂萃取分别获得雪菊石油醚萃取物(X-1)、乙酸乙酯萃取物(X-2)、正丁醇萃取物(X-3)和奥卡宁(X-2-1)。免疫磁珠法分离小鼠脾脏CD4^+T细胞,并以5×10^5个/孔的密度接种于96孔板,在加入刀豆蛋白A(ConA)的基础上分别给予不同浓度的萃取物和奥卡宁,共育48h后采用MTT法检测细胞的增殖情况。与此同时,进行细胞毒性实验,在不加入ConA的基础上分别给予不同浓度的萃取物和奥卡宁,共育48h后采用MTT法检测细胞的存活率。结果:雪菊乙酸乙酯萃取物(X-2)能显著抑制CD4^+T细胞的增殖,且呈剂量依赖性;X-2中的主要成分奥卡宁(X-2-1)同样表现为对CD4^+T细胞增殖的明显抑制,浓度为10μmol/L时细胞增殖率为(19.23±0.57)%,且在起效浓度下对正常细胞未表现出显著毒性。结论:雪菊乙醇提取物的乙酸乙酯萃取物及其含有的主要成分奥卡宁能显著抑制ConA介导的小鼠脾脏CD4^+T细胞的增殖。

Objective:To observe the effects of different extracts of Coreopsis tinctoria on proliferation of spleen-derived CD4+T cells of mouse in vitro.Methods:Sequoia petroleum ether extract(X-1),ethyl acetate extract(X-2),N-butanol extract(X-3)and okanin(X-2-1)were obtained by extracting with 95%ethanol and different polar solvents.Spleen-derived CD4^+T cells of mouse were isolated by immunomagnetic beads method and placed in 96-well plates at a cell density of 5×10^5 cells/well.Different concentrations of extracts and okanin were added to the basis of ConA,and MTT was used to detect the proliferation of cells after 48 h of co-culture.At the same time,cytotoxicity experiments were carried out,different concentrations of extracts and okanin were given without adding ConA.The survival rate of the cells was detected by MTT assay after 48 h of co-culture.Results:Ethyl acetate extract(X-2)inhibited the proliferation of CD4^+T cells in a concentration-dependent manner significantly.The main component of X-2,okanin,also showed the same effect,the cell proliferation rate was only(19.23±0.57)%at a concentration of 10μmol/L,and it showed no significant toxicity to normal cells at the effective concentration.Conclusion:Ethyl acetate extract of Coreopsis tinctoria and its main component,okanin,can significantly inhibit ConA-mediated proliferation of spleen-derived CD4^+T cells of mouse.