Relaxant effect and possible mechanism of 17-nor-subincanadine E in rabbit corpora cavernosa

Compounds with dual action on cyclic adenosine monophosphate （cAMP） and cyclic guanosine monophosphate （cGMP） may be a treatment option for erectile dysfunction, as they not only promote penile erection but also prevent the upregulation of phosphodiesterase-5. In this study, we examined the possible relaxant effect and mechanism of 17-nor-subincanadine E （SEC, 0.2-200 pmol I^-1）, a plant-derived alkaloid, in rabbit corpus cavernosum （RbCC） strips that had been precontracted by exposure to phenylephrine （10 pmol I^- 1） or a high concentration of K＋ （60 mmol I^- 1） in vitro. In addition to SEC＇s effect on cAMP and cGMP levels, electrical field stimulation （EFS） in phenylephrine-precontracted RbCC and calcium chloride （1-100 mmol I^-1） evoked responses in depolarized RbCC were analysed. SEC relaxed the phenylephrine-precontracted RbCCs in a concentration-dependent manner. Atropine, guanethidine and N-co-nitro-L-arginine methyl ester （L-NAME） did not have any effect on the relaxation of RBCCs. When 1H-1,2,4oxadiazole[4,3-a] quinoxalin-1-one （ODQ） was added, it effectively blocked the relaxant response of SEC. Although SEC enhanced the maximal relaxation produced by sodium nitroprusside （SNP） and forskolin in phenylephrine-precontracted cavernosal smooth muscle, it caused a decrease in the maximal contractile response induced by calcium chloride in depolarized RbCCs. The relaxant effect of SEC was paralleled by an increase in the tissue levels of the Cyclic nucleotides cAMP and cGMP. We conclude that SEC promotes the relaxation of RbCC, possibly favouring cAMP and cGMP accumulation and calcium blockade. This novel mechanism could be useful for patients who do not benefit from phosphodiesterase inhibitors and for those with endothelial and nitrergic dysfunction, such as patients with diabetes, hypertension and dyslipidaemias.

IL-1β activates p44/42 and p38 mitogen-activated protein kinases via different pathways in cat esophageal smooth muscle cells

AIM： To examine the pathway related to the IL-1β induced activation of mitogen-activated protein （MAP） kinases in cat esophageal smooth muscle cells. METHODS： Culture of the esophageal smooth muscle cells from cat was prepared. Specific inhibitors were treated before applying the IL-β3. Western blot analysis was performed to detect the expressions of COX, iNOS and MAP kinases. RESULTS： In the primary cultured cells, although IL-β3 failed to upregulate the COX and iNOS levels, the levels of the phosphorylated forms of 1344142 HAP kinase and p38 MAP kinase increased in both concentration- and time-dependent manner, of which the level of activation reached a maximum within 3 and 18 h, respectively. The pertussis toxin reduced the level of p44/42 MAP kinase phosphorylation. Tyrphostin 51 and genistein also inhibited this activation. Neomycin decreased the density of the p44/42 HAP kinase band to the basal level. Phosphokinase C （PKC） was found to play a mediating role in the IL-1β-induced p44/42 MAP kinase activity. In contrast, the activation of p38 MAP kinase was inhibited only by a pretreatment with forskolin, and was unaffected by the other compounds. CONCLUSION： Based on these results, IL-1β-induced p44/42 MAP kinase activation is mediated by the Gi protein, tyrosine kinase, phospholipase C （PLC） and PKC. The pathway for p38 MAP kinase phosphorylation is different from that of p44/42 MAP kinase, suggesting that it plays a different role in the cellular response to IL- 1β.