Chemical composition and antinociceptive and anti-inflammatory activity of the essential oil of Hyptis crenata Pohl ex Benth. from the Brazilian Amazon.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaf tea of Hyptis crenata has its practical use in the Brazilian Amazon for treating gastrointestinal and liver disorders, sweating induction, and as an anti-inflammatory. AIM OF THE STUDY: Evaluation of the chemical composition, acute oral toxicity, and antinociceptive and anti-inflammatory activities of the H. crenata essential oil. MATERIAL AND METHODS: The essential oil was hydrodistilled and analyzed by GC and GC-MS. The antinociceptive action in mice was evaluated for the peripheral and central analgesic activity (abdominal contortion and hot plate tests), and the xylene-induced ear swelling was carried out for the nociception test. RESULTS: Oxygenated monoterpenes (53.0%) and monoterpene hydrocarbons (38.9%) predominated in the H. crenata oil, being 1,8-cineo1e (35.9%), α-pinene (20.8%), camphor (10.0%), and ß-pinene (7.3%) their primary constituents. The oral oil administration in the mice did not display changes in behavior patterns or animal mortality at 300 and 2000 mg/kg doses. The control group's biochemical parameters (ALP, AST, ALT) displayed a statistical difference from the treated group, unlike the renal parameters, which showed no variation between the groups. Oil reduced the abdominal contortions at doses of 100 (79.5%) and 300 mg/kg (44.4%), while with endodontacin, the dose was 5 mg/kg (75.2%). In addition, the oil could not decrease the paw licking/biting time at doses of 30, 100, and 300 mg/kg. However, it showed a significant antinociceptive effect on the second phase in the formalin test inhibiting licking time, with a reduction of 50.8% (30 mg/kg), 63.4% (100 mg/kg), 58.0% (300 mg/kg), and morphine (4 mg/kg, 78.3%). The oil administration produced significant inhibition of ear edema at all tested doses, with a better effect produced at 30 mg/kg (64.0% inhibition). CONCLUSION: The oil of Hyptis crenata, rich in 1,8-cineole, camphor, α-pinene, and ß-pinene, totaling 74%, displayed low acute toxicity and significant anti-inflammatory activity, with peripheral and no central antinociceptive action. Thus, these results show an actual perspective on using H. crenata oil in developing a phytotherapeutic product.

The Essential Oil of Hyptis crenata Inhibits the Increase in Secretion of Inflammatory Mediators.

Background: Hyptis crenata is a plant of great ethnopharmacological importance widely distributed in South American countries. In Northeast Brazil, teas or infusions of its aerial parts are used in folk medicine to treat several acute and chronic inflammatory diseases. In a previous work we have demonstrated that the essential oil of H. crenata (EOHc) has an antiedematogenic effect. The aim of this work was to evaluate the effect of EOHc on cytokines secretion and cellular infiltration. Methods: Peritonitis and paw edema models induced by carrageenan were used to determine leucocyte count, myeloperoxidase (MPO) activity, nitrite, and cytokines secretion. Results: EOHc (10−300 mg/kg) significantly inhibited leucocyte migration and reduced the neutrophil count (control: 1.46 × 103 ± 0.031 × 103/mL) of the total leucocytes population in extracellular exudate (control: 2.14 × 103 ± 0.149 × 103/mL) by 15.00%, 43.29%, 65.52%, and 72.83% for the doses of 10, 30, 100, and 300 mg/kg EOHc, respectively (EC50: 24.15 mg/kg). EOHc (100 mg/kg) inhibited the increase in myeloperoxidase activity and completely blocked the increase in nitrite concentration induced by carrageenan. EOHc markedly reduced the pro-inflammatory cytokines (IL-6, MCP-1, IFN-γ, TNF-α, and IL-12p70) and increased IL-10, an anti-inflammatory cytokine (compared to control group, p < 0.05). Conclusions: This study demonstrates that EOHc has a long-lasting anti-inflammatory effect mediated through interference on MPO activity, and nitrite, and cytokines secretion. This effect, coupled with low EOHc toxicity, as far as results obtained in mice could be translated to humans, suggests that EOHc has great potentiality as a therapeutic agent.

Hepatoprotective Effect of Essential Oils from Hyptis crenata in Sepsis-Induced Liver Dysfunction.

No specific therapeutics are available for the treatment of sepsis-induced liver dysfunction, a clinical complication strongly associated with the high mortality rate of septic patients. This study investigated the effect of the essential oil of Hyptis crenata (EOHc), a lamiaceae plant used to treat liver disturbances in Brazilian folk medicine, on liver function during early sepsis. Sepsis was induced by the cecal ligation and puncture (CLP) model. Rats were divided into four groups: Sham, Sham+EOHc, CLP, and CLP+EOHc. EOHc (300 mg/kg) was orally administered 12 and 24 h after surgery. The animals were sacrificed for blood collection and liver tissue samples 48 h after surgery. Hepatic function was evaluated by measuring serum bilirubin, alkaline phosphatase (ALP), aspartate aminotransferase, and alanine aminotransferase (ALT) levels. The levels of malondialdehyde and the activity of superoxide dismutase, catalase, and GSH peroxidase (GSH-Px) were measured for assessment of oxidative stress. Liver morphology was analyzed by hematoxylin and eosin staining. EOHc normalized serum ALP, ALT, and bilirubin levels and inhibited morphological changes. In addition, we observed that EOHc inhibited elevation in hepatic lipid peroxidation and reduction of the glutathione peroxidase activity induced by sepsis. Our data show that EOHc plays a protective effect against liver injury induced by sepsis.

The essential oil of Hyptis crenata Pohl ex Benth. presents an antiedematogenic effect in mice

Hyptis crenata, commonly known as "salva-do-Marajó", "hortelã-do-campo", and "hortelãzinha", is used in folk medicine in Northeast Brazil as tea or infusion to treat inflammatory diseases. Due to the pharmacological efficacy and the low toxicity of the essential oil of Hyptis crenata (EOHc), we decided to investigate the EOHc antiedematogenic effect in experimental models of inflammation. EOHc was administrated orally at doses of 10-300 mg/kg to male Swiss albino mice. Paw edema was induced by subcutaneous injection in the right hind paw of inflammatory stimuli (carrageenan, dextran, histamine, serotonin, and bradykinin) 60 min after administration of EOHc. EOHc significantly inhibited the induced edema. The inhibitory effect of EOHc on dextran-induced edema extended throughout the experimental time. For the 30, 100, and 300 mg/kg doses of EOHc, the inhibition was of 40.28±1.70, 51.18±2.69, and 59.24±2.13%, respectively. The EOHc inhibitory effect on carrageenan-induced edema started at 10 mg/kg at the second hour (h) and was maintained throughout the observation period. At 30, 100, and 300 mg/kg doses the inhibition started earlier, from 30 min. At the edema peak of 180 min, 56, 76, and 82% inhibition was observed for 30, 100, and 300 mg/kg doses, respectively. Additionally, the effect of EOHc on carrageenan-induced paw edema was influenced by the time of administration. The EOHc also inhibited myeloperoxidase activity. In conclusion, the EOHc showed a potent effect, both preventing and reversing the edema, consistent with its anti-inflammatory use in folk medicine.

Gastroprotective effects of the essential oil of Hyptis crenata Pohl ex Benth. on gastric ulcer models.

ETHNOPHARMACOLOGICAL RELEVANCE: Hyptis crenata Pohl ex Benth (Lamiaceae), popularly known as "hortelã-brava" or "hortelã do campo", is widely distributed in the northeast of Brazil. In Brazil, the leaves of this plant have been used in traditional medicine for the treatment of gastrointestinal disturbances, including gastric ulcers. In an attempt to experimentally validate this claimed antiulcerogenic activity, the gastroprotective effects of the essential oil extracted from the leaves of the Hyptis crenata Pohl ex Benth (EOHc) were evaluated in recognized gastric ulcer models in mice. MATERIALS AND METHODS: EOHc was obtained by hydrodistillation and analyzed using gas chromatography-mass spectrometry (GC-MS). Swiss male mice (25-30g) were used for the studies. The gastric ulcers were induced by oral administration of absolute ethanol or indomethacin 45min after oral pretreatment with EOHc, vehicle and positive control drugs. One hour after the ulcerative challenges, the stomachs were removed and the area of the lesions was measured. The volume, pH and total acidity of the gastric secretions were determined using the pylorus ligature model. The gastrointestinal motility was measured using gastric emptying and intestinal transit. The ethanol-induced gastric mucus depletion and lipid peroxidation were also analyzed. RESULTS: Our findings are as follows: A significant inhibition of gastric lesions induced by absolute ethanol was observed in the mice pre-treated with EOHc, at a dose of 30 and 100 and 300mg/kg (5.56±1.51, 2.88±0.82 and 1.71±0.54mm(2), respectively) compared to control group (118.03±35.4mm(2)). Also, EOHc (300mg/kg) produced a gastroprotective effect against the gastric lesions induced by indomethacin (16.07±4.68mm(2)) compared to control group (38.64±6.1mm(2)). EOHc pretreatment produced a reduction in the ethanol-induced lipid peroxidation from 3.9±0.22 to 2.4±0.1µmol/mg tissue (EOHc-300mg/kg and control group, respectively). We also observed that EOHc pretreatment decreased the gastric emptying, but did not alter the intestinal transit ratio, ethanol-induced depletion of the gastric wall mucus or secretion parameters (volume, pH and [H(+)]). CONCLUSION: Our data indicate that EOHc exerts a gastroprotective effect, indicated by its significant inhibition of gastric lesions in ethanol- and indomethacin-induced ulcer models, which may be associated with its accelerating effect on gastric emptying and reduction in oxidative damages. Our data suggest a potential therapeutic application for EOHc in the treatment of gastric ulcers.

Antioxidant capacity and biological activity of essential oil and methanol extract of Hyptis crenata Pohl ex Benth / Capacidade antioxidante e atividade biológica do óleo essencial e extrato metanólico de Hyptis crenata Pohl ex Benth

O óleo essencial das folhas e ramos finos frescos e secos de Hyptis crenata forneceu os seguintes rendimentos, 1,4 por cento e 0,9 por cento. Os constituintes voláteis principais foram ±-pineno (22,0 por cento; 19,5 por cento), 1,8-cineol (17,6 por cento; 23,2 por cento), ²-pineno (17,0 por cento: 13,8 por cento), cânfora (4,7 por cento; 11,6 por cento), limoneno (5,4 por cento; 4,4 por cento) e ³-terpineno (3,5 por cento; 2,4 por cento), totalizando mais de 70 por cento nos óleos. A atividade de seqüestro do radical DPPH para o extrato metanólico (CE50, 16,7 + 0,4 µg/mL) foi comparável ao do BHT (19,8 ± 0,5 µg/mL) mostrando uma significante atividade antioxidante. Os óleos apresentaram baixa atividade. O teor de fenólicos totais (TP, 373,0 + 15,9 mg GAE/g) e equivalente trolox (TEAC, 226,8 + 0,5 mg TE/g) confirmaram a atividade antioxidante do extrato metanólico, que pode ser atribuída à presença de compostos fenólicos polares. No teste com larvas de camarão as concentrações letais para o óleo e extrato metanólico foram 6,7 + 0,2 µg/mL e 13,0 + 3,7 µg/mL, respectivamente, fornecendo importante evidência de suas atividade biológicas.

The essential oils of fresh and dried leaves and fine stems of Hyptis crenata furnished the following yields: 1.4 percent and 0.9 percent. The main volatile constituents were ±-pinene (22.0 percent; 19.5 percent), 1,8-cineole (17.6 percent; 23.2 percent), ²-pinene (17.0 percent; 13.8 percent), camphor (4.7 percent; 11.6 percent), limonene (5.4 percent; 4.4 percent) and ³-terpinene (3.5 percent; 2.4 percent), totalizing more than 70 percent in the oils. The DPPH radical scavenging activity (EC50, 16.7 + 0.4 µg/mL) of the methanol extract was comparable to BHT (19.8 ± 0.5 µg/mL) showing a significant antioxidant activity. The oils showed low activities. The amount of total phenolics (TP, 373.0 + 15.9 mg GAE/g) and trolox equivalent (TEAC, 226.8 + 0.5 mg TE/g) confirmed the antioxidant activity of the methanol extract that can be attributed to the presence of polar phenolic compounds. In the brine shrimp bioassay the lethal concentrations (LC50) for the oil and methanol extract were 6.7 + 0.2 µg/mL and 13.0 + 3.7 µg/mL, respectively, providing important evidence of their biological activities.