RESUMO
ETHNOPHARMACOLOGY RELEVANCE: Agastache mexicana is a popular plant of great demand in folk medicine, essentially due to its calming properties and for alleviating arthritic, muscular and abdominal pain. Despite its spectrum for pain relief, pharmacological studies of its bioactive constituents have been barely investigated. AIM OF THE STUDY: To evaluate protective properties of the A. mexicana and bioactive compounds improving pathological gastrointestinal conditions in rodents. MATERIAL AND METHODS: Different doses of the essential oil of A. mexicana ssp. mexicana and ssp. xolocotziana (30-562.2 mg/kg, i.p.) and individual monoterpenes (3-300 mg/kg, i.p.) were evaluated in an abdominal pain model. The most active monoterpene limonene and sulfasalazine (reference drug, 100 mg/kg, p.o.) were also evaluated in the oxazolone-induced colitis model using an oral gavage, where some inflammatory cytokines were analyzed by enzyme-linked immunosorbent assays. Finally, colonic histological assessment and gastroprotection in the absolute ethanol-induced ulcer model were explored. RESULTS: Our results demonstrated that the essential oil of both subspecies produced a significant reduction in the abdominal writhes, where monoterpenes limonene and pulegone were partially responsible bioactive metabolites. Limonene showed the major antinociceptive efficacy in the writhing test. It also significantly decreased hyperalgesia, pathological biomarkers, and colonic inflammatory cytokines in the oxazolone-induced colitis model, as well as prevention in gastric damage. CONCLUSIONS: Present results provide scientific evidence to reinforce the use of A. mexicana in the traditional medicine for gastrointestinal conditions, mainly related to pain and inflammation, demonstrating the potential of monoterpenes as natural products in the therapeutics of gastrointestinal affections such as ulcer, colitis, and abdominal pain.
Assuntos
Agastache/química , Analgésicos/farmacologia , Monoterpenos/farmacologia , Óleos Voláteis/farmacologia , Analgésicos/química , Analgésicos/isolamento & purificação , Animais , Colite Ulcerativa/tratamento farmacológico , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Fármacos Gastrointestinais/administração & dosagem , Fármacos Gastrointestinais/isolamento & purificação , Fármacos Gastrointestinais/farmacologia , Limoneno/administração & dosagem , Limoneno/isolamento & purificação , Limoneno/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Monoterpenos/química , Monoterpenos/isolamento & purificação , Óleos Voláteis/química , Óleos Voláteis/isolamento & purificação , Dor/tratamento farmacológico , Ratos , Ratos Wistar , Sulfassalazina/farmacologiaRESUMO
Parasitic diseases caused by gastrointestinal nematodes (GIN) are responsible for a major impact on ruminant welfare. Although the available anthelmintics have a safe margin of toxicity to the animals, their indiscriminate use has increased the selection of resistant parasite populations. In this scenario, essential oils (EO) stand out as a promising ecofriendly therapeutic alternative against GIN. The objective of this work was to determine the effect of the EO of Mentha villosa Hubs (MVEO) collected in 2017 and 2018, M. x piperita (MPEO) and their main components, carvone and limonene, against the third stage larvae (L3) of Haemonchus spp. and Trichostrongylus spp. The solutions, including in nanoemulsion preparations, were tested in a range of concentrations using the larval migration inhibition test (LMIT). The EO and carvone were also tested in combination with nitroxynil (NTX) to determine their effect as drug enhancers (additive or synergy). MVEO/2017, MVEO/2018, MPEO and carvone showed 70.6 (73.4â¯mg/mL), 86.3 (74.9â¯mL/mL), 95.5 (143.6â¯mg/mL), and 88.2 % (38.3â¯mg/mL) efficacy against L3, respectively. Carvone alone had approximately a 3-fold higher efficacy when compared to its concentration in each EO: 68.8 % in MVEO/2017 and 83.9 % in MVEO/2018. Limonene did not show any significant effect on inhibiting L3 migration. The combination of MPEO and NTX, and carvone and NTX showed a statistically significantly (Pâ¯<⯠0.05) synergic and additive effect, respectively, when compared to the isolated treatment. The nanoemulsion of MVEO/2017 at 0.367 mg/mL, inhibited L3 migration by 83.1 %, demonstrating to be highly effective (concentration ratio of 1:0.004), when compared to the MVEO/2017 (70.6 % at 73.4 mg/mL) extraction. The in vitro data from the combination of MPEO or carvone plus NTX suggest that these products can be considered for in vivo experiments against the most important GIN of ruminants as drug enhancers, possibly reducing the final concentration of NTX`. The efficacy of carvone was higher (EC50â¯=â¯1.96â¯mg/mL) than its expected efficacy, based on its concentrations on both EO. Therefore, this component does not need the entire EO composition to exert its L3 motility action. The remarkable efficacy demonstrated by the MVEO/2017/nanoemulsion (EC50â¯=â¯0.10â¯mg/mL), supports its potential to be a candidate to the next-generation therapy to alleviate clinical parasite infections and combat GIN resistant populations.
Assuntos
Anti-Helmínticos/farmacologia , Mentha/química , Nematoides/efeitos dos fármacos , Compostos Fitoquímicos/farmacologia , Óleos de Plantas/farmacologia , Animais , Anti-Helmínticos/administração & dosagem , Anti-Helmínticos/química , Monoterpenos Cicloexânicos/administração & dosagem , Monoterpenos Cicloexânicos/química , Monoterpenos Cicloexânicos/farmacologia , Sinergismo Farmacológico , Quimioterapia Combinada , Limoneno/administração & dosagem , Limoneno/química , Limoneno/farmacologia , Infecções por Nematoides/parasitologia , Infecções por Nematoides/veterinária , Nitroxinila/administração & dosagem , Nitroxinila/química , Nitroxinila/farmacologia , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Compostos Fitoquímicos/química , Óleos de Plantas/química , Ovinos , Doenças dos Ovinos/parasitologiaRESUMO
The aim of this study is to investigate the effects of limonene, alone or associated with therapeutic ultrasound, on oxidative stress following skeletal muscle injury. Thirty male Wistar rats were divided into 5 groups: CTR-control, MI-muscle injury without treatment, TPU-therapeutic pulsed ultrasound alone, TPU + LIM-phonophoresis with 5% limonene, and LIM-5% limonene applied topically. Muscle injury was induced by a mechanical abrupt impact over gastrocnemius muscle. The animals were treated in the following intervals: 2, 12, 24, 48, 72, and 96 h after injury. Blood and gastrocnemius samples were collected 98 h after lesion for data analysis. Creatine kinase (CK) and lactate dehydrogenase (LDH) activity, lipid peroxidation (TBARS) levels, catalase (CAT), and superoxide dismutase (SOD) activity were assessed. CK (p = 0.01), SOD activity (p < 0.01), and TBARS levels (p < 0.01) were increased after injury. There was no effect on LDH levels in any group. Phonophoresis (TABRS p < 0.01; SOD p = 0.01), TPU alone (TBARS p < 0.01; SOD p = 0.01), and LIM alone (TBARS p < 0.01; SOD p < 0.01) reduced TBARS levels and SOD activity after muscle injury. There was no change for CAT activity after injury. Only phonophoresis reduced CK activity after injury (p < 0.01). There was no difference between phonophoresis, TPU alone and LIM alone groups for TBARS, SOD, CAT, and LDH. Limonene alone and TPU alone were effective in reducing oxidative stress parameters after skeletal muscle injury. Only phonophoresis decreased CK activity. Skeletal muscle injury increases reactive oxidative species (ROS) levels and muscle proteins activity as creatine kinase (CK) and lactate dehydrogenase (LDH). Five percent limonene, alone or associated with therapeutic pulsed ultrasound, exhibited reduction of CK, superoxide dismutase (SOD) and catalase (CAT) activity, and lipid peroxidation markers (TBARS). Graphical abstract.
Assuntos
Antioxidantes/administração & dosagem , Limoneno/administração & dosagem , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/lesões , Estresse Oxidativo/efeitos dos fármacos , Fonoforese/métodos , Administração Tópica , Animais , Limoneno/metabolismo , Masculino , Músculo Esquelético/metabolismo , Estresse Oxidativo/fisiologia , Ratos , Ratos WistarRESUMO
BACKGROUND: In an increasing search for natural products that may heal the ulcers and avoid its recurrence, limonene appears as a promising candidate. HYPOTHESIS/PURPOSE: The present study aimed to investigate the protective effect of limonene in ethanol-induced gastric ulcers, in addition, to investigate the involvement of antioxidant and anti-inflammatory activities, besides the modulation of gene expression. STUDY DESIGN: Male Wistar rats were orally treated with vehicle (8% tween 80), carbenoxolone (100 mg/kg) or limonene (25, 50 or 100 mg/kg) and then orally received ethanol to induce gastric ulcers formation. METHODS: The activity of myeloperoxidase (MPO) was measured. Levels of glutathione (GSH) and activities of glutathione peroxidase (GPx), glutathione reductase (GR) and superoxide dismutase (SOD) were measured. We investigated the anti-inflammatory effect of limonene measuring the levels of pro-inflammatory cytokines tumor necrosis factor-a (TNF-a), interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and anti-inflammatory cytokine interleukin-10 (IL-10) by ELISA. Additionally, we investigate through real-time PCR (qPCR) the gene expression of nuclear factor-kappa B (Nf-κb), Gpx, Il-1ß, Mpo, and Il-10. RESULTS: Our results showed that limonene 50 mg/kg was the lowest effective dose, offering 93% of reduction in gastric ulcer area compared with the vehicle. There was an increase in mucus production and higher preservation of gastric mucosa integrity after treatment with limonene.There was a reduction in the MPO activity, a biomarker of neutrophils infiltration, and an increase in GPx activity, suggesting an antioxidant effect. Limonene displayed anti-inflammatory activity through decreasing the levels of TNF-a, IL-6, and IL-1ß and increasing the level of IL-10. Limonene could down-regulate the expression of Nf-κb, Il-1ß, and Mpo and up-regulate the expression of Gpx. CONCLUSION: Our results demonstrate that oral treatment with limonene exerts gastroprotection through local mucosal defense mechanisms, such as increasing the mucus production, modulation of the oxidative stress and inflammatory response and inhibition of Nf-κb expression.
Assuntos
Limoneno/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Úlcera Gástrica/prevenção & controle , Administração Oral , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/farmacologia , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Etanol/efeitos adversos , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/patologia , Gastrite/induzido quimicamente , Gastrite/tratamento farmacológico , Regulação da Expressão Gênica/efeitos dos fármacos , Glutationa/metabolismo , Limoneno/administração & dosagem , Masculino , Peroxidase/metabolismo , Ratos Wistar , Úlcera Gástrica/induzido quimicamente , Úlcera Gástrica/genética , Superóxido Dismutase/metabolismoRESUMO
This study was conducted to evaluate the effect of different concentrations of Limoneno (S)-(-) (50 µM,100 µM and 150 µM) in supplementing the diluter of bulls freezing semen. Thirty-six ejaculated from fourCurraleiro-Pé-Duro bulls were used for cryopreservation. The cryopreserved spermatozoa were submitted to postthawmotility sperm of computer assisted analysis (CASA) to evaluate the characteristics of spermatic kinetics.Observed that The different concentrations of limonene (S) - (-) did not affect the parameters of kinetics spermaticexcept for linearity (LIN). The addition of 150 μM limonene (S) - (-) significantly increased (P <0.05) the LINcompared to the control. The results obtained in the present study allow us to conclude that the supplementationof limonene (S) - (-) in cryopreservation bovine semen diluent did not interfere in kinetics sperm.
Assuntos
Masculino , Animais , Bovinos , Bovinos/embriologia , Criopreservação/métodos , Criopreservação/veterinária , Limoneno/administração & dosagem , Limoneno/análiseRESUMO
This study was conducted to evaluate the effect of different concentrations of Limoneno (S)-(-) (50 µM,100 µM and 150 µM) in supplementing the diluter of bulls freezing semen. Thirty-six ejaculated from fourCurraleiro-Pé-Duro bulls were used for cryopreservation. The cryopreserved spermatozoa were submitted to postthawmotility sperm of computer assisted analysis (CASA) to evaluate the characteristics of spermatic kinetics.Observed that The different concentrations of limonene (S) - (-) did not affect the parameters of kinetics spermaticexcept for linearity (LIN). The addition of 150 μM limonene (S) - (-) significantly increased (P <0.05) the LINcompared to the control. The results obtained in the present study allow us to conclude that the supplementationof limonene (S) - (-) in cryopreservation bovine semen diluent did not interfere in kinetics sperm.(AU)