温阳化浊通络方免疫抑制作用有效部位的筛选研究

Screening study on effective fractions of immune inhibition in Wenyang Huazhuo Tongluo Formula

目的:分离并筛选温阳化浊通络方治疗系统性硬化病免疫抑制作用的有效部位。方法:制备温阳化浊通络方水溶性部位、乙醚部位、乙酸乙酯部位、正丁醇部位、乙醇部位等不同溶剂提取物。在体内研究中,采用小鼠迟发型超敏反应(DTH)模型,观察温阳化浊通络方不同提取部位对细胞免疫和体液免疫的影响;体外研究检测温阳化浊通络方不同提取部位对Con A及LPS诱导的小鼠T、B淋巴细胞增殖的影响。结果:温阳化浊通络方水溶性部位和正丁醇部位能够明显减轻DTH小鼠耳肿胀、降低小鼠的脾指数和胸腺指数(P<0.01)。同时温阳化浊通络方水溶性部位和正丁醇部位不同剂量组均可抑制T、B淋巴细胞增殖,且呈一定剂量依赖性。结论:温阳化浊通络方治疗系统性硬化病免疫抑制作用的有效部位可能是水溶性部位和正丁醇部位。

Objective： To separate and screen the active fractions of immune inhibition from Wenyang Huazhuo Tongluo Formula （WYHZTLF） used for treatingsystemic sclerosis. Methods： Different extractfractions of WYHZTLF were prepared, including water-soluble fraction, ethereal fraction, ethyl acetate fraction, n-butyl alcohol fraction and ethanol fraction. In vivo experiment, mouse model of delayed-type hypersensitivity （DTH） was used to observe the influence of different extractfractions of WYHZTLF on cellular immunity and humoral immunity. In vitro experiment, the influence of different extractfractions of WYHZTLF on proliferation of T and B lymphocytes of mice induced by ConA and LPS was observed. Results： The water-soluble and n-butyl alcohol fractions of WYHZTLF could relieve the ear swelling and decrease spleen and thymus indexes of DHT mouse models （P〈0.01）, and different dosage of water-soluble and n-butyl alcohol fractions of WYHZTLF also could inhibit the proliferation of T and B lymphocytes simultaneously, which presented a dose-dependent characteristic. Conclusion： The water- soluble and n-butyl alcohol fractionsmight be confirmed as the effective fractions of immune inhibition in Wenyang Huazhuo Tongluo Formula for systemic sclerosis.