绒白乳菇中提取的乳菇菌素-D免疫抑制作用的研究

研究从绒白乳菇菌中分离出的一种新型倍半萜类化合物-乳菇菌素D的免疫抑制作用及机制。运用体外培养法取BALB/C小鼠脾细胞,研究乳菇菌素D对静止脾细胞、Con A(刀豆蛋白A)和LPS(脂多糖)刺激后的脾细胞(含B淋巴细胞和T淋巴细胞)增殖的抑制作用;运用MTT法经酶标仪检测吸光度,并计算刺激指数(stimulating index,SI);体外培养小鼠脾细胞,运用ELISA检测法观察乳菇菌素D对脾脏中T淋巴细胞分泌IL-2的抑制作用。结果显示该化合物对静止的脾细胞无抑制作用;对Con A和LPS刺激后的脾细胞增殖有显著抑制作用(P0.05),最大浓度药物组刺激指数SI低至0.28;高浓度的乳菇菌素D对Con A刺激的小鼠脾细胞产生的IL-2,具有显著抑制作用(P0.05)。结果显示乳菇菌素D通过抑制抗原刺激后的T淋巴细胞和B淋巴细胞的增殖、抑制其分泌细胞因子,从而发挥免疫抑制作用。研究结果将为开发以乳菇菌素D为母体、经结构修饰、安全低毒的免疫抑制剂提供理论基础。

Study on immunosuppressive effects of Mitissimol D isolated from Lactarius Vellereus

To study the immunosuppressive effects and the mechanism of a novel sesquiterpene (Mitissimol D) extracted from Lactarius Vellereus. The spleen cells from BALB/C mice in vitro culture were used to investigate the proliferation inhibition of Mitissimol D which to initial spleen cells, ConA and LPS-stimulated splenocytes (including B lymphocytes and T lymphocytes);the absorbance was detected by MTT assay with microplate reader and the stimulus index (SI) was calculatedS; the spleen cells of mice were cultured in vitro and the inhibitory effect of Mitissimol D on secretion of IL-2 from T lymphocytes of spleen was observed by ELISA. The results showed that the compound has no inhibitory effect on resting spleen cells ;but it significantly inhibited the spleen cells proliferation which were stimulated by ConA and LPS(P < 0.05), and the stimulus index of the drug group of the maximum concentration was as low as 0.28; Mitissimol D of high concentration had a significant inhibitory effect on the IL-2 secretion which produced by ConA-stimulated mouse spleen cells(P<0.05). Conclusion:The immunosuppressive effects of Mitissimol D was achieved by inhibiting the secretion of cytokines and inhibiting the proliferation of T lymphocytes and B lymphocytes which were stimulated by antigen. The results of this study would provide a theoretical basis for the development of safe and low toxicity immunosuppressive agents with mactosporin D as the parent and structural modification.