Screening Active Components Of Inhibiting Delayed Type Hypersensitivity From Yu-ping-feng Powder By Splenocyte Extracting Conjugated With HPLC (SE-HPLC)

Yu-ping-feng powder (YPS), a famous formula of traditional Chinese medicine, was widely used in treating recurrent respiratory tract infection in clinic as well as remedying allergic diseases. There are increasing evidences show that the studies of effective components in YPS enhancing immunity are focused on polysaccharides. However, the bioactive components for treating allergic diseases were undefined. This work was carried out to study the active ingredients against delayed type hypersensitivity (DTH) by using method of splenocyte extracting conjugated with HPLC (SE-HPLC).Among seven different extracts from YPS, ethanol extract of YPS were found out to inhibit the swelling of the auricle significantly on the model of mice allergic contact dermatitis (ACD) in vivo and proliferation of T lymphocyte and secretion of IFN-γinduced by ConA in vitro. Then ethanol extract were chosen to be the principal objects with splenocytes as the solid-phase extractor researching by SE-HPLC. Two compounds combining with splenocytes were detected by SE-HPLC and they were identified as PAN and Conjugated Linoleic acid (CLA) respectively. Non-sigificative changes in mice T lymphocyte proliferation were observed after treating with CLA. Whereas T lymphocyte proliferation and IFN-γsecretion levels induced by ConA were inhibited by PAN in vitro. Mice were administered with PAN during the induction phase of ACD. The extent of ear swelling was depressed conspicuously and the local pathological changes were improved obviously.Different from dexamethasone, no significant inhibition on immune organs in mice was observed after treated with PAN.The results suggest DTH was amended conspicuously after treating with PAN which combining with splenocytes screened from YPS by the method of SE-HPLC. These findings may help to evaluate SE-HPLC as a rapid and efficient method in screening active ingredients from complex system of traditional Chinese medicine.