The effects of five alkaloids from Bulbus Fritillariae on the concentration of cAMP in HEK cells transfected with muscarinic M(2) receptor plasmid.
The aim of this study was to investigate the effects of five alkaloids, namely verticine, verticinone, imperialine, imperialine-3beta-D-glucoside, and puqietinone, purified from Bulbus Fritillariae and used as an antitussive drug in traditional Chinese medicine, on their antimuscarinic M(2) function and the cAMP level of HEK cells transfected with muscarinic M(2) receptor plasmid. By transfecting the HEK cells with the method of calcium phosphate co-precipitation and screening with G418, the cells stably expressing M(2) receptor were identified. The expression of M(2) receptor in HEK cells was confirmed by both RT-PCR and western blot. The cAMP level in the treated cells was analyzed with RIA method ((125)I-cAMP KIT). And the results suggested that the five alkaloids could significantly elevate the cAMP concentration in the HEK cells transfected with muscarinic M(2) receptor plasmid (p < 0.01).
Zhou
Ying
Y
Key Laboratory of Modern Chinese Medicines, Ministry of Education of PRC and Department of Pharmacognosy, China Pharmaceutical University, Nanjing 210009, China.
Ji
Hui
H
Lin
Bao-Qin
BQ
Jiang
Yan
Y
Li
Ping
P
D000470:Alkaloids / Q000737:chemistry / Q000494:pharmacology*; D015153:Blotting, Western; D002460:Cell Line; D002595:Cevanes / Q000737:chemistry / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D031410:Fritillaria / Q000737:chemistry*; D005960:Glucosides / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D015394:Molecular Structure; D018517:Plant Roots / Q000737:chemistry; D043585:Receptor, Muscarinic M2 / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D000470:Alkaloids; D002595:Cevanes; D005960:Glucosides; D043585:Receptor, Muscarinic M2; C068448:imperialine-beta-glucoside; C014242:verticine; C069537:sinpeinine A; D000242:Cyclic AMP; C047331:peiminine