ABSTRACT
BACKGROUND: Based on the traditional application of traditional Chinese Medicines (TCMs), Ephedra Herba (EH) is used to cure cold fever by inducing sweating, whereas Ephedra Radix (ER) is used to treat hyperhidrosis. Although they come from the same plant, Ephedra sinica Stapf, but have play opposing roles in clinical applications. EH is known to contain ephedrine alkaloids, which is the driver of the physiological changes in sweating, heart rate and blood pressure. However, the active pharmacological ingredients (APIs) of ER and the mechanisms by which it restricts sweating remain unknown. PURPOSE: The current work aims to discover the hidroschesis APIs from ER, as well as to establish its action mechanism. METHODS: UPLC-Q/TOF-MS, PCA, and heat map were utilized for identifying the differences between EH and ER. HPLC integrated with a ß2-adrenoceptor (ß2-AR) activity luciferase reporter assay system was used to screen active inhibitors; molecular docking and a series of biological assays centered on ß2-AR-related signaling pathways were evaluated to understand the roles of APIs. RESULTS: The opposite effect on sweating of EH and ER can be attributed to the APIs of amphetamine-type alkaloids and flavonoid derivatives. Mahuannin B is an effective anti-hydrotic agent, inhibiting the production of cAMP via suppression of adenylate cyclase (AC) activity. CONCLUSION: The effects of EH and ER on sweat and ß2-AR-related signaling pathway are opposite due to different alkaloids and flavonoids of APIs in EH and ER. The present work not only sheds light on the hidroschesis action of mahuannin B, but also presents a potential target of AC in the treatment of hyperhidrosis.
Subject(s)
Adenylyl Cyclase Inhibitors/pharmacology , Alkaloids/pharmacology , Cyclic AMP/metabolism , Drugs, Chinese Herbal/pharmacology , Ephedra/chemistry , Flavonoids/pharmacology , Receptors, Adrenergic, beta-2/metabolism , Adenylyl Cyclase Inhibitors/chemistry , Adrenergic beta-2 Receptor Antagonists/chemistry , Adrenergic beta-2 Receptor Antagonists/pharmacology , Alkaloids/chemistry , Animals , Chromatography, High Pressure Liquid , Drug Evaluation, Preclinical/methods , Drugs, Chinese Herbal/chemistry , Ephedra sinica/chemistry , Ephedrine/pharmacology , Flavonoids/chemistry , Male , Mice , Molecular Docking Simulation , Receptors, Adrenergic , Signal Transduction/drug effects , Species Specificity , Sweating/drug effectsABSTRACT
Objective: To study the flavonoids from the roots of Ephedra sinica in Ephedra Tourn. ex L. Methods: The roots of E. sinica were extracted with 90% ethanol, and isolated and purified by silica gel, Sephadex LH-20, and RP-18 gel column chromatography. All the compounds were identified based on spectral analyses (including MS, 1H-NMR, and 13C-NMR). Results: Thirteen flavonoids were isolated from the roots of E. sinica and were characterized as ephedrannin A (1), mahuannin A (2), mahuannin B (3), mahuannin D (4), apigenin (5), kaempferol (6), quercetin (7), dihydroquercetin (8), 3′, 4′, 5, 7-tetrahydroxy flavanone (9), (+)-catechin (10), (-)-epi-catechin (11), afzelechin (12), and (-)-epi-afzelechin (13), respectively. Conclusion: Compounds 7-13 are obtained from this plant for the first time.
