The combination of rat mast cell and rabbit aortic smooth muscle is the simple bioassay for the screening of anti-allergic ingredient from methanolic extract of Corydalis tuber.
We have assessed the release of histamine from mast cells by smooth muscle contraction. 0.3 microg/ml compound 48/80 showed no effect on concentration-response relationship of histamine in rabbit aorta. Compound 48/80 induced release of histamine from rat mast cells. When aorta was stimulated by compound 48/80 in the presence of mast cells, contraction was evoked in concentration-dependent manner. This mast cell-dependent contraction was completely blocked by H1 receptor antagonist, 1 microM diphenhydramine. When mast cells was treated with compound 48/80 inhibitor benzalkonium chloride, mast cell-dependent contraction was inhibited, although benzalkonium chloride itself showed no effect on concentration-response relationship of histamine in rabbit aorta. At high concentration of 10 microg/ml, benzalkonium chloride itself evoked histamine release from mast cells and indeed inhibitory effect of 10 microg/ml benzalkonium chloride on mast cell-dependent contraction was lower than that of 3 microg/ml. We have applied this bioassay to search anti-allergic ingredient from a total methanolic extract of Corydalis tuber (Corydalis turtschaninovii BESSER forma yanhusuo Y. H. CHOU et C. C. HSU). Successively, we have isolated five fractions. The fractions I-IV are identified to be corybulbine (1), tetrahydropalmatine (2), corydaline (3) and yuanhunine (4), respectively. Main component of fraction V is the mixture of 3 and canadine (5). Fractions II and V significantly inhibited mast cell-dependent contraction in rabbit aorta as well as inhibited histamine release from rat mast cells. Furthermore, fractions I, III and V inhibited histamine-induced contraction in rabbit aorta at non-competitive manner. From these results, combination of rat mast cells and rabbit aorta is good bioassay to search the anti-allergic ingredient, and we have obtained effective fractions from Corydalis tuber using this assay.
Saito
Shin-Ya
SY
Department of Pharmaceutical Molecular Biology, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.
Tanaka
Masashi
M
Matsunaga
Kimihiro
K
Li
Yushan
Y
Ohizumi
Yasushi
Y
D000818:Animals; D018926:Anti-Allergic Agents / Q000494:pharmacology*; D001013:Aorta, Thoracic / Q000187:drug effects*; D001681:Biological Assay; D031311:Corydalis / Q000737:chemistry*; D006636:Histamine Release; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D008407:Mast Cells / Q000187:drug effects*; D000432:Methanol / Q000737:chemistry; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D010936:Plant Extracts / Q000494:pharmacology*; D011817:Rabbits; D051381:Rats
D018926:Anti-Allergic Agents; D010936:Plant Extracts; D000432:Methanol