José Henrique Leal-Cardoso

Antihypertensive effects of the essential oil of Alpinia zerumbet and its main constituent, terpinen-4-ol, in DOCA-salt hypertensive conscious rats

The present study investigated the hypotensive responses to intravenous (i.v.) treatment with the essential oil of Alpinia zerumbet (EOAZ) and its main constituent, terpinen‐4‐ol (Trp‐4‐ol), in the experimental model of deoxycorticosterone‐acetate (DOCA)‐salt hypertensive rat.

Vasorelaxant effects of the monoterpenic phenol isomers, carvacrol and thymol, on rat isolated aorta

Various essential oils are rich in carvacrol, a monoterpenic phenol isomeric with thymol. This study was undertaken to assess the vasorelaxant effects of thymol and carvacrol in rat isolated aorta and the putative mechanisms underlying these effects. Thymol and carvacrol produced a concentration‐dependent relaxation on the aortic ring preparations pre‐contracted using KCl (IC50 value of 64.40 ± 4.41 and 78.80 ± 11.91 μm, respectively) or using phenylephrine (PHE, 0.1 μm) (IC50 value of 106.40 ± 11.37 and 145.40 ± 6.07 μm, respectively) and inhibited the concentration‐response curves of aortic rings to PHE or KCl. In Ca2+‐free medium with ethylene glycol‐bis(2‐aminoethylether)‐N,N,N′,N′‐tetraacetic acid (2 mm), thymol and carvacrol both at 1000 μm completely abolished the phasic component of PHE‐induced endothelium‐containing ring contractions. At 400 μm, thymol and carvacrol significantly reduced the CaCl2‐induced contractions in Ca2+‐free medium. Furthermore, both thymol and carvacrol (300 and 1000 μm) significantly reduced the contraction evoked by phorbol dibutyrate (1 μm), an activator of protein kinase C. Magnitude of this inhibitory effect was enhanced in the presence of the Ca2+ pump inhibitor, thapsigargin (1 μm). At 1000 μm, neither thymol nor carvacrol altered the resting potential of vascular smooth muscle cells. In conclusion, thymol and carvacrol induced an endothelium‐independent relaxation in rat isolated aorta, an effect that seems mediated through some mechanisms probably involving a transduction pathway between Ca2+ release from sarcoplasmic reticulum and/or regulation of the Ca2+ sensitivity of the contractile system. Moreover, it’s conceivable that thymol and carvacrol, at low concentrations, block the Ca2+ influx through the membrane.

Myorelaxant and antispasmodic effects of the essential oil of Alpinia speciosa on rat ileum

The effects of the essential oil of Alpinia speciosa Schum (EOAS) on rat ileum were studied. EOAS (0.1–600 µg/mL) reversibly relaxed ileal basal tonus. Submaximal contractions induced by 60 mM KCl or acetylcholine were concentration dependently inhibited by EOAS with similar IC50 values (∼ 44 and 48 µg/mL, respectively). These results show that EOAS possesses both relaxant and antispasmodic actions in the ileum. Copyright

Relaxant effects of the essential oil of Ocimum gratissimum on isolated ileum of the guinea pig

The effects of the essential oil of Ocimum gratissimum L. (Labiatae) (EOOG) on guinea pig ileum were studied. EOOG (0.1-1000 microg/ml) reversibly and concentration-dependently relaxed the basal tone of the ileum and reversed the tonic contractions induced by 60 mM KCl and 10 microM acetylcholine, with IC(50) values of 23.8+/-5.2, 18.6+/-4.0 and 70.0+/-4.6 microg/ml, respectively. Our results show that EOOG exerts relaxant effects on intestinal smooth muscle, consistent with the popular use of the plant to treat gastrointestinal disorders.

Intestinal myorelaxant and antispasmodic effects of the essential oil of Croton nepetaefolius and its constituents cineole, methyl-eugenol and terpineol

The effects of the essential oil of Croton nepetaefolius (EOCN), a medicinal plant from the north‐east of Brazil, and its constituents cineole, methyl‐eugenol and terpineol, were studied on intestinal motility in vivo and on in vitro mechanical activity of intestinal smooth muscle. In mice, EOCN (10–100 mg/kg body weight, intragastrically) increased the intestinal transit of charcoal marker delivered to the stomach. This was also observed in animals pretreated with castor oil. In segments of guinea‐pig ileum and cardial, pyloral and ileo‐caecal sphincters, EOCN preferentially decreased basal tonus compared with the amplitude of spontaneous contractions with EC50 values in the range 0.9 – 16 and 8 – 150 μg/mL respectively. In ileum, EOCN, cineole, methyl‐eugenol and terpineol decreased tonus with EC50 values of 16, 322, 9 and 71 μg/mL, respectively, and blocked 60 mM [K+]‐induced contraction with IC50 values of 18, 419, 12 and 95 μg/mL.

In vivo anti-inflammatory action of eugenol on lipopolysaccharide-induced lung injury

Eugenol, a methoxyphenol component of clove oil, suppresses cyclooxygenase-2 expression, while eugenol dimers prevent nuclear factor-kappaB (NF-kappaB) activation and inflammatory cytokine expression in lipopolysaccharide-stimulated macrophages. Our aim was to examine the in vivo anti-inflammatory effects of eugenol. BALB/c mice were divided into four groups. Mice received saline [0.05 ml intratracheally (it), control (Ctrl) and eugenol (Eug) groups] or Escherichia coli LPS (10 microg it, LPS and LPSEug groups). After 6 h, mice received saline (0.2 ml ip, Ctrl and LPS groups) or eugenol (160 mg/kg ip, Eug and LPSEug groups). Twenty-four hours after LPS injection, pulmonary resistive (DeltaP1) and viscoelastic (DeltaP2) pressures, static elastance (E(st)), and viscoelastic component of elastance (DeltaE) were measured. Lungs were prepared for histology. In parallel mice, bronchoalveolar lavage fluid was collected 24 h after LPS injection. TNF-alpha was determined by ELISA. Lung tissue expression of NF-kappaB was determined by EMSA. DeltaP1, DeltaP2, E(st), and DeltaE were significantly higher in the LPS group than in the other groups. LPS mice also showed significantly more alveolar collapse, collagen fibers, and neutrophil influx and higher TNF-alpha levels and NF-kappaB expression than the other groups. Eugenol treatment reduced LPS-induced lung inflammation, improving lung function. Our results suggest that eugenol exhibits in vivo anti-inflammatory action in LPS-induced lung injury.

Linalool blocks excitability in peripheral nerves and voltage-dependent Na+ current in dissociated dorsal root ganglia neurons

Linalool is a terpene that occurs as a major constituent of essential oils of many plants of widespread distribution. It possesses several biological and pharmacological activities, including depressant effects on the central nervous system and olfactory receptors. The present study investigated whether linalool affects the excitability of peripheral components of the somatic sensory system. We used sciatic nerve and preparations of intact and dissociated neurons of dorsal root ganglion for extracellular, intracellular and patch-clamp recordings. Linalool concentration-dependently (0.3-2.0mM) and reversibly blocked the excitability of the sciatic nerve. It inhibited peak-to-peak amplitude of the compound action potential (IC(50) was 0.78+/-0.04 mM). At 0.8mM, it reversibly increased rheobase and chronaxy (from 3.2+/-0.1 V and 52.4+/-4.1 micros to 4.2+/-0.3 V and 71.2+/-5.5 micros (n=5), respectively) and inhibited with greater pharmacological potency the amplitude of the compound action potential components corresponding to axons with slower velocity of conduction. In a similar concentration range (0.1-6mM), linalool concentration-dependently and reversibly blocked the generation of action potentials of intact dorsal root ganglion neurons without alteration of resting membrane potential and input resistance, and inhibited the voltage-gated Na(+) current of dissociated dorsal root ganglion neurons. In conclusion, we demonstrated that linalool acts on the somatic sensory system with local anesthetic properties, since it blocked the action potential by acting on voltage-dependent Na(+) channels. This finding is important in showing the potential usefulness of linalool as a pharmacotherapeutic agent.

EFFECTS OF PIPERITENONE OXIDE ON THE INTESTINAL SMOOTH MUSCLE OF THE GUINEA PIG

We investigated the effects of piperitenone oxide (PO), a major constituent of the essential oil of Mentha x villosa, on the guinea pig ileum. PO (30 to 740 micrograms/ml) relaxed basal tonus without significantly altering the resting membrane potential. In addition, PO relaxed preparations precontracted with either 60 mM K+ or 5 mM tetraethylammonium in a concentration-dependent manner. At concentrations from 0.1 to 10 micrograms/ml PO potentiated acetylcholine-induced contractions, while higher concentrations (> 30 micrograms/ml) blocked this response. These higher PO concentrations also inhibited contractions induced by 60 mM K+. PO also blocked the components of acetylcholine contraction which are not sensitive to nifedipine or to solutions with nominal zero Ca2+ and EGTA. These results show that PO is a relaxant of intestinal smooth muscle and suggest that this activity may be mediated at least in part by an intracellular effect.

Antinociceptive effects of the essential oil of Croton zehntneri in mice

Croton zehntneri is an aromatic plant native to Northeastern Brazil, where it is often used in folk medicine. In the present study the antinociceptive effects of the essential oil of Croton zehntneri (EOCz) were evaluated in mice. EOCz administered orally at doses of 100 and 300 mg/kg reduced paw licking time in the second phase of the formalin test from the control value of 41.61 +/- 8.62 to 12.01 +/- 7.97 and 6.57 +/- 3.42 s, respectively. During the first phase of the formalin test only 300 mg/kg induced a significant alteration (from 58.2 +/- 7.02, control, to 28.7 +/- 4.73 s). The number of contortions in response to intraperitoneal injections of acetic acid did not differ significantly between controls (80.6 +/- 9.01) and experimental (300 mg/kg body weight) animals (89.1 +/- 9.53% of the control numbers; P > or =0.05, Student t-test). In the hot-plate test, EOCz at doses > or =100 mg/kg significantly increased the latency time with respect to controls (11.2 +/- 0.80). At 100 and 300 mg/kg this increase persisted for 180 and 240 min, respectively. The data show that EOCz is effective as an antinociceptive agent.

Effects of the essential oil of Croton zehntneri, and its constituent estragole on intestinal smooth muscle

The effects on isolated guinea‐pig ileum of the essential oil of Croton zehntneri (CZEO) and of its main constituent estragole (57% of CZEO by weight) were studied. CZEO and estragole (0.1–100 μg/mL) decreased the tonus in 56% and 61.5%, respectively, of the muscles. At concentrations above 10 μg/mL, they induced spontaneous rhythmic movements of small amplitude (less than 11% of the potassium contraction peak to peak). At concentrations from 1 to 100 μg/mL and with similar potencies, these agents blocked the contractions induced by acetylcholine, histamine and 50 mM K+ and caused relaxation of already established potassium contractures. Tested separately, CZEO, estragole and anethole (28% of CZEO by weight) blocked the contraction induced by Ca++ in the presence of 50 mM K+, but CZEO was more potent than estragole or anethole in blocking the Ca++‐induced contractions than those induced by K+. With large increases in the agonist concentration, the action of the oils on the contractions induced by Ca++ was reversible; however, their effect on contractions induced by histamine or ACh was not. The data show that the essential oil of Croton zehntneri has an effect on intestinal smooth muscle that is predominantly antispasmodic, and attributable in part to the effect of estragole, a major constituent.