RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Colocasia esculenta (CE) (L.) Schott is an annual herbaceous tropical plant from the family of Araceae which has been traditionally used for the healing of various ailments such as asthma, arthritis, internal hemorrhage, diarrhea, and neurological disorders. The plant is reported to have potential anti-microbial, anti-fungal, antimetastatic, anti-hepatotoxic, and anti-lipid peroxidative activities. AIM OF THE STUDY: The present study is designed to explore the potential anti-inflammatory property of Colocasia esculenta methanolic root extract (CEMRE) on carrageenan-induced rat paw edema and lipopolysaccharide (LPS) stimulated RAW264.7 cells. MATERIALS AND METHODS: Carrageenan-induced rat paw edema model was used to investigate the in vivo anti-inflammatory action of CEMRE. Adult male Wistar rats (180-220 g; n = 6) were pre-treated with CEMRE (100, 200, and 400 mg/kg BW) orally before 1 h of injection of 1% carrageenan. Indomethacin (10 mg/kg BW) was given orally as the standard drug. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), nitric oxide (NO), prostaglandinE2 (PGE2), and cytokines levels were measured. Liquid chromatography-mass spectrometry (LC-MS) was done to identify the phytoconstituents present in CEMRE. The inhibitory activity of CEMRE was investigated against cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in in vitro assessment of LPS-stimulated RAW264.7 cells. The RAW 264.7 cells were pre-treated with Indomethacin (5 µM and 10 µM) and CEMRE (17 µg/ml and 34 µg/ml) followed by induction of LPS (1 µg/ml) for 24 h. Docking analyses were also performed to explore the interaction of important phytoconstituents (Sinapic acid, Acetylsalicylic acid, L-fucose, Salicylic acid, Quinic acid, Zingerone, and Gingerol) of CEMRE with COX-2 and iNOS. RESULTS: Pre-treatment with CEMRE (400 mg/kg) could inhibit the paw inflammation significantly which was elevated due to carrageenan induction. The inhibition is comparable to that of the standard drug Indomethacin. The concentration of serum AST, ALT, ALP, NO, PGE2 and cytokines were also considerably lowered in the CEMRE-treated group as compared to the carrageenan-induced group. CEMRE (34 µg/ml) inhibited the LPS-stimulated relative expression of mRNA of COX-2 and iNOS and significantly reduced the expression of nitric oxide and prostaglandin E2. Docking analyses revealed promising interaction with low binding energies between Sinapic acid with both the target proteins COX-2 and iNOS. CONCLUSION: Collectively, our results suggested that CEMRE exhibited effective anti-inflammatory actions on carrageenan-induced rat paw edema and LPS-treated RAW 264.7 cells by reducing the in vivo paw edema inhibition, inhibiting the serum NO, PGE2, cytokines and also reduced the in vitro production of NO, PGE2 along with expressions of mRNA COX-2 and iNOS. Molecular docking demonstrated good binding affinities among the target proteins and ligand Sinapic acid. Thus the bioactive compound from CE need to be isolated and purified.
Assuntos
Anti-Inflamatórios , Colocasia , Animais , Ratos , Anti-Inflamatórios/farmacologia , Carragenina , Colocasia/química , Ciclo-Oxigenase 2/metabolismo , Citocinas/metabolismo , Dinoprostona/metabolismo , Edema/induzido quimicamente , Edema/tratamento farmacológico , Edema/patologia , Indometacina , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Lipopolissacarídeos , Simulação de Acoplamento Molecular , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Ratos Wistar , Células RAW 264.7 , CamundongosRESUMO
The Aquaporins (AQPs) could prove to be striking targets of inflammation. The aim of this study was to study the involvement of AQPs and explore the anti-inflammatory activity of Garcinia extract in LPS induced acute systemic inflammation in Wistar rats. Adult male Wistar rats (n = 6) were pretreated with Garcinia orally twice for 7 days, followed by a single intraperitoneal dose (5.5 mg/kgbw) of LPS. Serum ALT, AST, ALP, Creatinine, Urea and BUN, nitric oxide, prostaglandin, cytokine and chemokine levels were measured. LC-MS analysis of Garcinia was performed to identify the phytoconstituents present. The iNOS and COX enzyme activity were determined in the target tissues. qPCR analysis of inos, cox-2 and aqps was performed. Relative protein expression of AQPs was studied by Western blot analysis. Molecular docking studies were performed to study the interaction of garcinol and hydroxycitric acid, the two important phytoconstituents of Garcinia with AQP. The qPCR analysis showed tissue-specific up-regulation of aqp1, aqp3, aqp4 and aqp8 in LPS induced rats. Garcinia extract treatment effectively lowered the mRNA expression of these AQPs. Garcinia extract significantly inhibited the LPS-induced NO, prostaglandin, cytokine and chemokine production in serum and also decreased tissue-specific transcript level of inos and cox-2, thus suggesting the anti-inflammatory role of Garcinia. Also, docking studies revealed interactions of garcinol and hydroxycitric acid with AQP1, 3, 4 and 8. Therefore, the present study suggests the possible involvement of AQP1, 3, 4 and 8 in inflammation and the efficacy of Garcinia extract as an anti-inflammatory agent. Therefore, AQPs can act as prognostic markers of inflammation and can be targeted with Garcinia extract.
Assuntos
Anti-Inflamatórios/uso terapêutico , Aquaporinas/antagonistas & inibidores , Garcinia , Mediadores da Inflamação/antagonistas & inibidores , Lipopolissacarídeos/toxicidade , Extratos Vegetais/uso terapêutico , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Aquaporinas/biossíntese , Relação Dose-Resposta a Droga , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Mediadores da Inflamação/metabolismo , Masculino , Simulação de Acoplamento Molecular/métodos , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Estrutura Secundária de Proteína , Ratos , Ratos Wistar , Resultado do TratamentoRESUMO
Effective extraction of phyto-biomolecules insures retaining maximum functionality along with higher recovery. In this study, ultrasound-solvent assisted extraction (USAE) was employed for optimal extraction of phyto-biomolecules from Sesamum indicum (sesame) leaves using the approach of Response Surface Methodology (RSM). The optimized condition of 200 W power, 59% methanol concentration with 1:14 g/mL solid-liquid ratio and 15 min of extraction time yielded 367.39 ± 1.85 mg GAE/100 g of total phenolic content, 96.72 ± 3.27% of free radical scavenging activity and 81.20 ± 2.87% of iron chelating activity respectively. The extract consist of essential phytocomponents like gallic acid, chlorogenic acid, and quercetin with lipid peroxidation activities of >50% over incubation time of 48 h. Also, showed antimicrobial activity against various Gram's negative and positive food borne pathogens. The results of this study implied the importance of USAE for effective and optimum recovery of phyto-biomolecules from Sesame leaves with retained functional properties.
Assuntos
Fenóis/isolamento & purificação , Extratos Vegetais/química , Sesamum/química , Anti-Infecciosos , Sequestradores de Radicais Livres , Ácido Gálico/isolamento & purificação , Quelantes de Ferro , Peroxidação de Lipídeos , Fenóis/farmacologia , Extratos Vegetais/farmacologia , Folhas de Planta/química , Ondas UltrassônicasRESUMO
OBJECTIVE: The purpose of this study was to investigate and measure the variable morphologies of axis vertebrae and explore the clinical significance of variations as it may pertain to clinical palpation and diagnostic imaging. METHODS: The common variable morphologies in 100 specimens of intact dry adult axis vertebrae (Chinese) were investigated and measured. The frequencies in deviation of odontoid processes, deviation of spinous processes, and presence of bifid spinous processes were observed. The distances between the apices of transverse processes and inferior articular facets were also measured. RESULTS: Variable morphologies of C2 that we observed were deviation of odontoid processes (14 cases, 14.0%), deviation of spinous processes (3 cases, 3.0%), and bifid spinous processes (95 cases, 95.0%). Of the bifid spinous processes, 56 had a process on the left side equal to the right side, 21 were longer on the left, and 18 were longer on the right. The distances between apices of transverse processes and inferior articular facets in the left side of C2 were 17.67 +/- 2.47 mm, and that of the right side were 17.81 +/- 2.55 mm. CONCLUSIONS: Because variable morphology of the axis is common, congenital deviation of the odontoid process, deviation of the spinous process, and asymmetrical bifid spinous processes should be taken into account during clinical palpation and diagnostic imaging.
Assuntos
Vértebra Cervical Áxis/anatomia & histologia , Adulto , Povo Asiático , Diagnóstico por Imagem , Humanos , Técnicas In Vitro , Processo Odontoide/anormalidades , Processo Odontoide/anatomia & histologia , Processo Odontoide/patologia , Palpação , FotografaçãoRESUMO
OBJECTIVE: The purpose of this study was to investigate and measure morphological changes in the vertebral canal and its cast form at the level of the cervical spine in vitro during flexion, extension, and lateral bending in cadaver specimens. METHODS: The morphological changes of vertebral canal and its contents were investigated and measured during experimental flexion, extension, and lateral bending of the cervical spine with 10 fresh specimens (Chinese); the cross-sections and sagittal diameters were also measured by pouring liquid wax into the intervertebral canal. RESULTS: During lateral bending, the nucleus pulposus was pushed to the opposite side and the inferior cervical nerve roots of the opposite side were stretched. Cross-sectional diameter at the level of C6-7 during flexion was larger than that in lateral bending (P < .05). Comparing extension with flexion, we found that changes in all segments' areas were significant (P < .05). There was no significant difference in the sagittal diameter at any segment during all postures (P > .05). CONCLUSIONS: During lateral bending, the nucleus pulposus of neck were pushed into the opposite side, and inferior cervical nerve roots of the opposite side were stretched. The C5, C6, and C7 nerve roots appeared to undergo excessive stretch when an excessive lateral bending beyond the physiologic range was undergone. This study provides some additional evidence about the mechanics of cervical spine motion.
Assuntos
Vértebras Cervicais/anatomia & histologia , Postura , Canal Medular/anatomia & histologia , Anatomia Transversal , Cadáver , Vértebras Cervicais/inervação , Dura-Máter/anatomia & histologia , Humanos , Técnicas In Vitro , Ligamento Amarelo/anatomia & histologia , Masculino , Modelos Anatômicos , Parafina , Medula Espinal/anatomia & histologia , Raízes Nervosas Espinhais/anatomia & histologia , Estresse MecânicoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Xiao-Xu-Ming decoction (XXMD) has long been employed clinically to treat stroke in traditional Chinese Medicine. AIM OF THE STUDY: To investigate the neuroprotective effects of XXMD in vivo and in vitro stroke models and determine involved mechanisms. MATERIALS AND METHODS: Two models (four-vessel occlusion in adult Wistar rats and oxygen-glucose deprivation primary cultured neurons) were employed to mimic ischemia-reperfusion damage, in vivo and in vitro, respectively. The effects of XXMD were investigated with respect to neuronal damage, activity of caspase-3 and expression of Bcl-2 in CA1 region of hippocampus after ischemia. The cognitive ability was measured 7 days after ischemia/reperfusion by using Morris water maze. RESULTS: Oral administration of XXMD significantly increased the density of neurons that survived in the CA1 region of hippocampus on the 3rd and 7th day after transient global ischemia was induced in a dose-dependent manner. XXMD ameliorated severe deficiencies in spatial cognitive performance induced by transient global ischemia. Inhibition of caspase-3 activity and up-regulation of Bcl-2 expression were induced in the high dose of XXMD-treated rats after ischemia. In oxygen-glucose deprivation model, both XXMD extract and drug-containing serum prepared from blood of high dose of XXMD-treated rats inhibited apoptotic neuronal death at 24h after reoxygenation. CONCLUSIONS: Our results clearly demonstrated that XXMD is neuroprotective and appears to influence deleterious pathological processes that are activated after the onset of ischemia.