Mechanisms involved in the contractile responses induced by the hydroalcoholic extract of Phyllanthus urinaria on the guinea pig isolated trachea: evidence for participation of tachykinins and influx of extracellular Ca2+ sensitive to ruthenium red.
1. The hydroalcoholic extract (HE) of stems, leaves and roots from P. urinaria (Euphorbiaceae) (1-3000 micrograms/ml), caused graded contraction in guinea pig trachea (GPT), being more effective in preparations without epithelium. 2. Response to HE was slightly affected by tetrodotoxin (0.3 microM) and nicardipine (1 microM), but was unaffected by w-conotoxin, atropine, mepyramine or staurosporine (all 1 microM). Indomethacin (3 microM) greatly inhibited HE contraction, but MK 571 (leukotriene D4 and E4 antagonist) caused partial inhibition; L-655,240 (thromboxane A2 antagonist) and WEB 2086 (PAF antagonist) (all 1 microM) were ineffective. 3. Response to HE was markedly inhibited in a Ca(2+)-free solution and was partially affected in GPT desensitized to capsaicin (10 microM). 4. Capsazepine (capsaicin antagonist, 3 microM) antagonized the contraction from capsaicin, leaving the response to HE unaffected. In contrast, ruthenium red (an ionic channel antagonist coupled to vanilloid receptors of capsaicin) (0.1-3 microM) caused graded and equipotent noncompetitive inhibition of HE- and capsaicin-induced contractions, but had no effect on carbachol- and prostaglandin E2-mediated responses. 5. FK 888 and SR 48968 (NK1 and NK2 receptor antagonists, respectively) (both 1 microM) antagonized, through a competitive mechanism, the contraction from SP and [beta-ala8]NKA (4-10) respectively, but antagonized, through a noncompetitive mechanism, HE-mediated contraction. 6. We concluded that contraction to HE in GPT is modulated by the epithelium, depends on the release of a cyclo-oxygenase metabolite, and relies largely upon an extracellular Ca2+ influx that is highly sensitive to ruthenium red, but is insensitive to L and N-type of voltage-sensitive Ca2+ channel antagonists. In addition, NK1 and NK2 tachykinins, but not vanilloid receptors, play an important role in mediating its response.
Paulino
N
N
Department of Pharmacology, Universidade Federal de Santa Catarina, Brazil.
Cechinel Filho
V
V
Pizzolatti
M G
MG
Yunes
R A
RA
Calixto
J B
JB
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism; D005260:Female; D006168:Guinea Pigs; D066298:In Vitro Techniques; D020024:Leukotriene Antagonists; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects* / Q000378:metabolism; D064729:Neurokinin-1 Receptor Antagonists; D010936:Plant Extracts / Q000494:pharmacology; D010946:Plants, Medicinal / Q000737:chemistry*; D010972:Platelet Activating Factor / Q000037:antagonists & inhibitors; D011448:Prostaglandin Antagonists / Q000494:pharmacology; D018041:Receptors, Neurokinin-2 / Q000037:antagonists & inhibitors; D012430:Ruthenium Red / Q000494:pharmacology; D013373:Substance P / Q000037:antagonists & inhibitors; D015320:Tachykinins / Q000037:antagonists & inhibitors / Q000502:physiology*; D014132:Trachea / Q000187:drug effects* / Q000378:metabolism
D002121:Calcium Channel Blockers; D020024:Leukotriene Antagonists; D064729:Neurokinin-1 Receptor Antagonists; D010936:Plant Extracts; D010972:Platelet Activating Factor; D011448:Prostaglandin Antagonists; D018041:Receptors, Neurokinin-2; D015320:Tachykinins; D012430:Ruthenium Red; D013373:Substance P; D002118:Calcium