桂枝汤方剂不同提取部位免疫抑制活性的考察

Screening for immunosuppressive function on different fraction of Guizhi decoction

目的筛选桂枝汤方剂免疫抑制活性部位。方法处方药材分别水煎浓缩为水提部位,乙醇提取为乙醇提取部位。同时将乙醇部位分别用石油醚、氯仿、乙酸乙酯、正丁醇萃取得石油醚部位、氯仿部位、乙酸乙酯部和正丁醇部位。CCK-8法考察桂枝汤不同提取物对Con A激活T细胞和LPS激活B细胞增殖的抑制作用。同时用二硝基氯苯对昆明种小鼠皮肤致敏和诱发,制成迟发型超敏反应模型,观察桂枝汤方剂不同部位对迟发型超敏反应的干预作用。结果桂枝汤方剂水提物、乙醇提取物、石油醚萃取部位有显著免疫抑制功效,乙醇和石油醚提取物效果最佳。浓度200 mg·L-1时T细胞增殖抑制率分别为48.3%、45.9%,B细胞抑制率大于30%。乙醇提取物、石油醚萃取部位对迟发型超敏反应小鼠耳廓肿胀有显著抑制作用,对胸腺和脾脏无明显影响。结论桂枝汤方剂乙醇提取物和石油醚萃取物为免疫抑制活性部位,能显著抑制T细胞和B细胞的增殖,抑制小鼠耳廓肿胀,对正常细胞、胸腺、脾脏无毒性。

Objective To screen the immunosuppressive fraction of Guizhi decoction.Methods Medicinal materials of Guizhi decoc-tion was extracted by water and ethanol to get water extraction and ethanol extraction.Ethanol extraction was extracted with petroleum e-ther,chloroform,ethyl acetate,n-butanol to get petroleum ether fraction,chloroform fraction,ethyl acetate fraction,and n-butanol fraction respectively.CCK-8 method was used to detect the activity of different extracts on lymphocytes in the spleen of mice.ConA and LPS were added to the spleen cells of BALB/c mice to activate T cells and B cells respectively.Different extracts of Guizhi decoction were added to the activated spleen cells to investigate the immunosuppressive effect.The model of delayed-type hypersensitivity was estab-lished by sensitizing the skin of Kunming mice with DNCB.Effects of different fractions of Guizhi decoction were observed on delayed-type hypersensitivity mice.Results Water extraction,ethanol extraction and petroleum ether fraction had significant immunosuppres-sion.Ethanol extraction and petroleum ether fraction showed the greatest activities with inhibitory rates of T cells 48.3% and 45.9%, and B cells inhibitory rate was more than 30% at the concentration of 200 mg·L ^-1 .Ethanol extraction and petroleum ether fraction could alleviate obviously the degree of swollen-auricle of mice and without significant influence on spleen and thymus.Conclusions The ethanol extraction and petroleum ether fraction were the main immunosuppressive fraction,which could significantly inhibit the pro-liferation of T and B cells and alleviate the swollen-auricle of mice without toxicity to normal cells and spleen and thymus.