大补阴丸对自身免疫病模型小鼠免疫调节的血清药理学研究

A serum-pharmacological study on the immunoregulatory effects of DaBu Yin pills in autoimmune mice induced by Campylobacter jejuni

本文探讨大补阴丸（汤）（DBYW）试验血清对空肠弯曲菌（Campylobacter jejuni，CJ）致敏的自身免疫病模型小鼠（CJS135小鼠）T淋巴细胞分泌IFN-γ／IL-4活性的影响及其时间-效应关系。将健康雌性ICR小鼠64只随机分为正常对照组（N）、模型组（M）、模型对照组（MC）以及五个时相给药组（30min、1h、1.5h、2h、3h），共8组。除正常对照组外，其他各组均用空肠弯曲菌免疫小鼠制备自身免疫病模型。从造模第15天起，开始以大补阴丸（汤）（0．4g生药饮片／ml）给不同时相用药组小鼠灌胃，连续用药10d。并分别于第10次给药后30min、1h、1.5h、2h、3h取血，分离得试验血清；同时取N组、MC组血清，作对照。各组小鼠脾细胞经体外培养，检测DBYW试验血清对T淋巴细胞分泌IFN-γ／IL-4活性的影响，并绘出时-效关系变化曲线。实验结果显示，大补阴丸（汤）可明显降低实验小鼠体内异常升高的dsDNA、ssDNA抗体水平，改善其因自身免疫反应所致的肝肾病理损害，小鼠服药后1～1.5h两时相试验血清，可明显抑制小鼠T淋巴细胞分泌IFN-γ活性，升高细胞培养上清中IL-4水平，其免疫调节作用存在明显的时效关系。提示大补阴丸（汤）试验血清对异常免疫机能状态下的T淋巴细胞分泌IFN-γ／IL-4活性具有明显的免疫调节作用，且在服药后1～1．5h作用最明显。其对于自身免疫病小鼠的体液免疫和细胞免疫机能均存在双向调节作用。

To explore the immunoregulatory effect of Da-Bu-Yin Wan（DBYW） on cytokines of IFN-γ and IL-4 in autoimmune mice induced by Campylobacter jejuni （CJ） with a method of serum-pharmacology study, healthy female ICR mice were divided into 8 groups at random： normal control group（N）, model group（M）, model control group（MC）, and groups with administration of medicinas at different times（30 min. 1 h. 1.5 h. 2 h. 3 h）. The mice were induced by Campylobacterjejuni with complete Freund＇s adjuvant as autoimmune animal model except N group. From the fifteenth day after primary immunization, the administration groups were given DBYW through gastric route in a dosage of 0.4 g/ml. At the same time, MC group were given physiological saline instead. All the administration lasted for 10 days. After the last administration, blood samples were taken after 30 min, 1 h, 1.5 h, 2 h and 3 h of medication respectively to cultivate T lymphocytes which were taken from M group. The concentrations of IFN-γ and IL-4 in culture supernantants were measured by enzyme-linked immunosorbent assay（ELISA） after 48 h. The drug efficacy was evaluated by marking the curve of time-effect relationship in the chart with time along the horizontal axis versus inhibition rate of pathological changes along the vertical axis. It was found that the concentrations of IFN-γ was significantly lower in cultures with the drug-containing serum taken at 1 - 1.5 h after the treatment than that in cultures with the MC control serum（P〈0.01）. Only the serum taken at 1.5 h was found to be able to enhance the production of IL-4（P〈0.05）. These results indicate that the drug-containing serum in mice taken at the time of 1- 1.5 hrs after ingestion of DBYW can regulate the production of IFN-γ and IL-4 in autoimmune mice induced by Campylobacter jejuni.