OBJECTIVE To extract the -SH active fractions (SHF) from Bubali Cornu (water buffalo horn) and evaluate its antipyretic activity.
METHODS SHF was extracted from Bubali Cornu by SDS-DTT, and the content of native thiols (-SH) was determined by Ellman reagent method. SHF was identified based on nano LC-MS/MS technology. Evaluation of antipyretic activity of SHF was based on LPS-induced fever rat model. The levels of PGE2, IL-1β, and TNF-α in plasma as well as the levels of cAMP, PGE2, and TNF-α in the hypothalamus were measured by ELISA kits. An untargeted metabolomics approach was used to further investigate the intervention of SHF on plasma metabolites in febrile rats.
RESULTS SDS-DTT could effectively extract SHF from Bubali Cornu, in which the main components were type Ⅰ, type Ⅱ keratins and keratin-associated proteins, which were rich in Cys, and the ratio of -SH to protein in SHF was increased about 20 times more than that of traditional decoction. SHF could significantly decrease (P < 0.01) the body temperature which lasted for 4.5 hours. SHF could also significantly decrease the levels of PGE2, IL-1β, TNF-α and cAMP in plasma and hypothalamic. A total of 137 potentially differential metabolites were identified from plasma samples of the control and model groups, of which 31 metabolites could be dialed back after SHF administration, including lysophosphatidic acid, phosphatidylinositol, phosphatidic acid, triglycerides, phosphatidylcholine and so on, which were mainly involved in the glycerophospholipid metabolic pathway.
CONCLUSION SHF has precise antipyretic effect, and the dosage of 1/10 of the aqueous extract can show its comparable antipyretic effect, which provides the direction and basis for the basic research on the antipyretic efficacy of Bubali Cornu.