麻黄-甘草药对对小鼠脾细胞的增殖以及Th1/Th2平衡的影响
Effect of Ephedra Sinica-Glycyrrhiza Uralensis Herbal Compatibility on the Proliferation of Splenocytes and Th1/Th2 Balance in Mice
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摘要: 目的 研究麻黄-甘草药对对小鼠脾细胞的增殖以及在刀豆蛋白A刺激下对Th1/Th2平衡的影响，探讨其对免疫功能的作用与机制。方法 利用MTT法考察不同质量浓度的药对及单体对脾淋巴细胞增殖的影响；药对干预刀豆蛋白A(5 mg/L)建立的淋巴细胞Th1/Th2平衡模型，qPCR法检测药对及单体对刀豆蛋白A刺激条件下脾淋巴细胞Th1型常见标志物IFN-γ、T-bet及Th2型常见标志物IL-4、GATA3基因的表达，ELISA法检测IL-4与IFN-γ蛋白水平。结果 25~100 mg/L浓度的麻黄-甘草药对对静息状态以及刀豆蛋白A诱导的脾细胞有促增殖作用；麻黄碱、伪麻黄碱对静息状态以及刀豆蛋白A诱导的脾细胞有促增殖作用；甘草苷与甘草酸在0.01~10 μmol/L浓度范围内对脾细胞增殖未见明显影响。在调节Th1/Th2平衡方面，麻黄-甘草药对可以显著抑制IL-4与GATA3的表达，而对IFN-γ与T-bet的表达未见明显影响；其中麻黄碱能显著上调IFN-γ与T-bet的表达，伪麻黄碱既能抑制IL-4与GATA3的表达，也能抑制IFN-γ与T-bet的表达；甘草酸能抑制IL-4与GATA3的表达，同时能上调IFN-γ与T-bet的表达（P<0.01）；甘草苷对上述基因未见明显影响；药对及单体均能抑制IL-4的蛋白水平，其中以药对、麻黄碱与伪麻黄碱作用更明显（P<0.05～0.01）；药对与甘草苷能抑制IFN-γ的蛋白水平，麻黄碱、伪麻黄碱及甘草酸能提高IFN-γ的蛋白水平，其中甘草酸的作用最明显（P<0.05）。结论 麻黄-甘草药对及其单体能对静息状态与活化状态的脾细胞产生促增殖作用，并能通过调节Th1/Th2平衡影响机体的免疫功能。Abstract: OBJECTIVE To investigate the effects of Ephedra sinica-Glycyrrhiza uralensis herbal compatibility on the proliferation of splenocytes and the balance of Th1/Th2 under the stimulation of concanavalin A(ConA)， and explore its effect on immune function. METHODS Effect of different concentration of couplet medicines and monomers on splenic lymphocyte proliferation in mice was determined by MTT assay; the Th1/Th2 balance model of lymphocyte was established by ConA (5 mg/L); PCR was used to detect the mRNA expression of IFN-γ，T-bet，IL-4 and GATA3 in splenic lymphocytes of mice under ConA stimulation; ELISA was used to detect the protein expression of IL-4 and IFN-γ. RESULTS 25-100 mg/L of couplet medicines showed proliferative activities of splenocytes in both rest state and ConA-induced state; ephedrine and pseudoephedrine at a dose of 0.1-10 mg/L showed a proliferative effect in both rest state and ConA-induced state; glycyrrhizin and glycyrrhizin showed no proliferative effect on splenocytes. Ephedra sinica-Glycyrrhiza uralensis herbal compatibility significantly inhibited the mRNA expression of IL-4 and GATA3 (P<0.01)， while displayed no effect on the mRNA expression of IFN-γ and T-bet. Ephedrine significantly up-regulated the mRNA expression of IFN-γ and T-bet (P<0.01)， but had no effect on the mRNA expression of IL-4 and GATA3; pseudoephedrine inhibited the mRNA expression of IL-4， GATA3， IFN-γ and T-bet (P<0.01); glycyrrhizic acid inhibited the expression of mRNA IL-4 and GATA3 (P<0.01)， but up-regulated the expression of IFN-γ and T-bet (P<0.01); glycyrrhizin showed no effect on Th1/Th2 balance in splenic lymphocytes. Couplet medicine and its monomers all inhibited the level of IL-4 protein. Couplet medicine and glycyrrhizin inhibited the level of IFN-γ; ephedrine， pseudoephedrine and glycyrrhizic acid increased the level of IFN-γ， and glycyrrhizic acid showed the most obvious effect (P<0.05). CONCLUSION Couplet medicine and its monomer can promote proliferation of spleen cells on rest state and ConA-induced splenocytes， and improve the immune function through the regulation of Th1/Th2 balance.
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