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目的:探索玉屏风散(YPFS)诱导初期给药对Th2(Ⅱ型辅助性T细胞)型接触性超敏反应的效应,并利用HPLCMS(液质联用)法测定玉屏风散含药血清中的成分。方法:Th2型接触性超敏反应造模采用Balb/c小鼠第1,2天用1.5%的异硫氰酸荧光素(FITC)溶液致敏腹部皮肤,第6天用0.6%的异硫氰酸荧光素(FITC)溶液攻击小鼠耳部皮肤,24 h后测定小鼠耳厚和耳重。致敏前给予地塞米松ip剂量为0.67 mg·kg-1,YPFS(3.25,6.5 g·kg-1,ig),毛蕊异黄酮(0.5,5,10 mg·kg-1,ip),芒柄花素(0.5,5,10 mg·kg-1,ip)2 d,并在致敏第1~3天继续给药3 d,第4天停止给药(诱导初期给药)。含药血清检测采用ICR小鼠每天给予YPFS醇提物,连续给药3 d,空白对照组给予等体积的生理盐水。最后1次给药后1 h取血处死动物。血清经过乙酸乙酯萃取处理后,利用HPLC-MS法进行测定。结果:与正常组相比,模型组小鼠耳肿胀度及耳重显著增加(P<0.01)。玉屏风散醇提物3.25,6.5 g·kg-1诱导初期给药能显著抑制小鼠耳肿胀度(P<0.01或P<0.05),减轻过敏性炎症。玉屏风散醇提物含药血清经HPLC-MS测定出3个主要成分,经鉴定分别为升麻素、毛蕊异黄酮、芒柄花素。毛蕊异黄酮和芒柄花素5,10 mg·kg-1诱导初期给药能显著抑制Th2型接触性超敏反应模型小鼠耳肿胀度,芒柄花素10 mg·kg-1诱导初期给药显著降低耳重。结论:玉屏风散诱导初期给药能显著抑制FITC诱导的小鼠Th2型变应性接触性皮炎的耳肿胀度,含药血清测得的入血成分经过药效学实验验证,对Th2型过敏反应有抑制作用,表明血清中存在抗过敏作用的活性成分。
OBJECTIVE: To explore the effect of YPFS-induced initial administration on Th2 (type II helper T cell) contact hypersensitivity and use HPLCMS (liquid chromatography-mass spectrometry) to determine the content of Yupingfengsan The ingredients. Methods: Th2-type hypersensitivity was induced in Balb / c mice by sensitizing abdominal skin with 1.5% fluorescein isothiocyanate (FITC) solution on day 1 and day 2. On the 6th day, The solution of fluorescein cyanide (FITC) attacked the ear skin of mice and the ear thickness and ear weight were measured 24 h later. The dexamethasone ip dose was 0.67 mg · kg-1, YPFS (3.25,6.5 g · kg-1, ig), calyciflorin (0.5, 5 and 10 mg · kg- (0.5, 5, 10 mg · kg-1, ip) for 2 days and continued for 3 days on days 1 to 3 of sensitization and stopped on the 4th day (induction of initial administration). Drug-containing serum was detected using ICR mice daily YPFS alcohol extract, continuous administration of 3 d, blank control group given an equal volume of saline. The animals were sacrificed 1 h after the last administration. Serum was extracted with ethyl acetate and assayed by HPLC-MS. Results: Compared with the normal group, the auricular swelling and ear weight of mice in the model group increased significantly (P <0.01). Yupingfengsan ethanol extract 3.25,6.5 g · kg-1 initial administration can significantly inhibit mice ear swelling (P <0.01 or P <0.05), reduce allergic inflammation. Yupingfeng San alcohol extract containing serum by HPLC-MS determination of the three main components, were identified as cimicifuga, calycosin, calyx. The initial administration of 5 and 10 mg · kg -1 of calycosin and flavonoids could significantly inhibit the eardrum swelling of mice with Th2-type hypersensitivity reaction, and the initial administration of 10 mg · kg-1 Significantly reduce ear weight. Conclusion: The initial administration of Yupingfeng Powder can significantly inhibit ear swelling induced by FITC-induced mouse allergic contact dermatitis of Th2 type. The blood components measured by the drug-containing serum are validated by pharmacodynamic experiments and are sensitive to Th2 allergy The reaction has an inhibitory effect, indicating that there is an anti-allergic active ingredient in the serum.