Recent studies revealed an involvement of RhoA/Rho-kinase signaling in the agonist-induced Ca2+ sensitization of bronchial smooth muscle contraction, and the pathway has now been proposed as a new target for the treatment of airway obstructive diseases, such as asthma. On the other hand, volatile anesthetics such as isoflurane are traditionally used to treat status asthmaticus. In the present study, the effect of inhibition of Rho-kinase on the isoflurane-induced relaxation of bronchial smooth muscle was investigated. Smooth muscle strips of intrapulmonary bronchi obtained from Wistar rats were used. Application of isoflurane (0.5-4.0%, generated by a calibrated vaporizer) to the acetylcholine (30 μM)-precontracted rat bronchial smooth muscles caused a concentration-dependent relaxation. Interestingly, the isoflurane-induced relaxation was significantly augmented by the pretreatment with subthreshold concentration of Y-27632, a Rho-kinase inhibitor. Thus, the combined use of Y-27632 and isoflurane might be useful for treatment of severe airway blockade, such as status asthmaticus.
The leaf of Psidium guajava Linn. (family: Myrtaceae) is used traditionally in African folk medicine to manage, control and/or treat a plethora of human ailments, including diarrhoea. In this study, we examined the antidiarrhoeal activity of Psidium guajava leaf aqueous extract (PGE) on experimentally-induced diarrhoea in rodents. PGE (50-400 mg/kg p.o.) produced dose-dependent and significant (P<0.05-0.01) protection of rats and mice against castor oil-induced diarrhoea, inhibited intestinal transit, and delayed gastric emptying. Like atropine (1 mg/kg, p.o.), PGE produced dose-dependent and significant (P<0.05-0.01) antimotility effect, and caused dose-related inhibition of castor oil-induced enteropooling in the animals. Like loperamide (10 mg/kg, p.o.), PGE dose-dependently and significantly (P<0.05-0.01) delayed the onset of castor oil-induced diarrhoea, decreased the frequency of defaecation, and reduced the severity of diarrhoea in the rodents. Compared with control animals, PGE dose-dependently and significantly (P<0.05-0.01) decreased the volume of castor oil-induced intestinal fluid secretion, and reduced the number, weight and wetness of faecal droppings. PGE also produced concentration-related and significant (P<0.05-0.01) inhibitions of the spontaneous, rhythmic, pendular contractions of the rabbit isolated duodenum. The findings of this study indicate that PGE possesses antidiarrhoeal activity, and thus lend pharmacological credence to the suggested folkloric use of the plant as a natural remedy for the treatment, management and/or control of diarrhoea in some rural communities of southern Africa.
Mefloquine, an antimalarial drug, has been reported to block exogenously transfected gap junctions composed of either Cx36 or Cx50 more potently than those composed of other connexins. Using the conventional whole-cell clamp technique, we investigated the effects of mefloquine on intercellular electrical coupling in vascular endothelial cells of guinea-pig mesenteric arteries, where expressions of Cx40 and Cx43 have been identified. Mefloquine (50 μM) almost abolished the current required to impose a 10 mV command step, leaving only rapid capacitive currents and some sustained currents after about 3 min. The relaxation of the capacitive current could be well fitted with a single exponential function. The effect of mefloquine was reversible and the time course of the current induced by the voltage step gradually changed back after mefloquine was removed. The mean input resistance and capacitance in the presence of mefloquine were 323 MΩ and 10.1 pF, respectively. While intercellular electrical coupling was well blocked by mefloquine (50 μM), the membrane hyperpolarized from -24.0 to -32.5 mV. The results indicate that mefloquine effectively blocks gap junctions without producing major side effects in vascular endothelial cells and that this compound is a useful tool in the investigation of gap junctions.
In this study, we examined the effects of Psidium guajava Linn. leaf aqueous extract (PGE) on isolated, spontaneously-contracting portal veins, as well as on endothelium-intact and endothelium-denuded descending thoracic aortic ring preparations of healthy, normotensive rats. Graded concentrations of PGE (0.25-4.0 mg/ml) caused concentration-dependent, initial brief but significant (P<0.05) rises of the basal tones and amplitudes of pendular, rhythmic contractions, followed by secondary pronounced, longer-lasting and significant (P<0.05-0.001) inhibitions of contractile amplitudes of the isolated portal veins. Relatively low concentrations of PGE (<1.0 mg/ml) always contracted freshly-mounted, naïve, endothelium-intact aortic ring preparations. However, relatively high concentrations of PGE (1.0-4.0 mg/ml) always produced initial brief contractions/augmentations of noradrenaline (NA, 10-7M)-induced contractions of endothelium-intact and endothelium-denuded aortic ring preparations, followed by secondary, pronounced relaxations of the aortic ring muscles. Moreover, relatively high concentrations of PGE (1.0-4.0 mg/kg) always relaxed NA-induced contractions of the aortic ring preparations in a concentration-related manner. The arterial-relaxing effects of PGE were more pronounced in endothelium-intact aortic rings than in endothelium-denuded aortic ring preparations. The relaxant effects of PGE on endothelium-intact aortic rings were only partially inhibited by NG-nitro-L-arginine methyl ester (L-NAME, 100 μM), a nitric oxide synthase inhibitor, suggesting that the vasorelaxant effect of PGE on aortic rings is probably mediated via both endothelium-derived relaxing factor (EDRF)-dependent and EDRF-independent mechanisms. Taken together, the findings of this study indicate that PGE possesses a biphasic effect on rat isolated vascular smooth muscles.
The frequency of slow waves recorded from circular muscle bundles with attached longitudinal muscle (intact muscle) was compared with that of slow potentials recorded from isolated circular muscle bundles (isolated muscle) from the guinea-pig stomach. In intact muscle preparations, slow waves were generated in the corpus, antrum and pylorus with a higher frequency in the corpus (about 5 min-1) than the other regions (about 2 min-1 in antrum, about 1.5 min-1 in pylorus). The resting potential amplitude was graded across the stomach, at about -50 mV in the fundus, -60 mV in the corpus, -65 mV in the antrum and -70 mV in the pylorus. A similar distribution of resting membrane potential and slow potential frequency was also observed in isolated muscle bundles from the different regions. Caffeine (1 mM) abolished slow waves in some corpus preparations and inhibited the 2nd component of slow waves in the antrum and pylorus, and also abolished slow potentials in isolated muscle preparations from any region of the stomach. This suggests that myenteric interstitial cells of Cajal (ICC-MY) are heterogeneously distributed in the stomach (pylorus, antrum and part of the corpus regions), with a homogeneous distribution of muscular interstitial cells of Cajal (ICC-IM) within the circular muscle bundles. The frequency of slow potentials in smooth muscle isolated from any region of the stomach changed linearly in response to membrane potential changes produced by either current injection or high potassium solutions. The frequency of slow potentials after setting the membrane potential at -60 mV was larger in the corpus than the antrum, suggesting that the high frequency discharge of corpus muscle is produced by the low membrane potential and additional unidentified factors. We suggest that the regional difference in slow wave discharge is produced mainly by ICC-IM, and the role of ICC-MY may be little, if any.