RESUMO
AIM OF THE STUDY: The study was performed to understand the detailed mechanism of diabetic wound healing by bilirubin-deferoxamine (DFO) combination on topical application. MATERIALS AND METHODS: There were two study groups, control, and treatment. The granulation tissues collected on different days (3, 7, 14, and 19) were studied in detail for inflammatory mediators, angiogenesis markers, epithelialization, and oxidative stress parameters. RESULTS: A significant increase in wound contraction percentage was observed from day 7 in the bilirubin-DFO treatment group. The combinatorial treatment significantly reduced tumour necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1ß), and enhanced IL-10 levels. Upregulated mRNAs of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1 alpha (HIF-1 α) along with CD31 immunohistochemistry showed the pro-angiogenesis potential of the combination. Hematoxylin and Eosin (H and E) staining and Masson's trichrome staining showed reduced inflammatory cell infiltration, enhanced fibroblast proliferation, well-organized collagen fibers, and the development of new blood vessels. Collagen deposition is further supported by immunohistochemistry studies and Masson's trichrome staining. Bilirubin-DFO combination also reduced lipid peroxidation and elevated antioxidative enzymes. CONCLUSION: Topical application of bilirubin-DFO showed immense potential in augmenting skin wound regeneration in diabetes by upregulating the antioxidant status as well as increasing angiogenesis, collagen deposition, and modulating cytokines.
Assuntos
Desferroxamina , Diabetes Mellitus Experimental , Animais , Antioxidantes , Bilirrubina/metabolismo , Colágeno/farmacologia , Colágeno/uso terapêutico , Desferroxamina/metabolismo , Desferroxamina/farmacologia , Desferroxamina/uso terapêutico , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Estresse Oxidativo , Ratos , Pele , Fator A de Crescimento do Endotélio Vascular , CicatrizaçãoRESUMO
S-[3-carbamoylsulfanyl-2-(dimethylamino)propyl] carbamothioate (Cartap) (CAS number: 15263-52-2) is a synthetic insecticide of thiocarbamates group that is extensively used in field of agriculture for controlling of several pests like rice stem borer, leaf folder pests in paddy field and diamond back moth, aphids in cabbage and cauliflower crops. Cartap, as a pesticide has not been investigated yet for its effect on vital organs and biochemical stress in vivo and the present study was undertaken to evaluate the same in Swiss albino mice. For this purpose male mice were given three different dose levels of cartap, i.e. 5 mg/kg, 7.5 mg/kg and 15 mg/kg body weight respectively, for 28 days orally. Water was used as vehicle to dissolve cartap. Oral administration of cartap caused significant increase in serum biomarkers, tissue oxidants and decrease in antioxidants along with histopathological findings in liver, kidney and brain tissues. Thus, present study showed that in vivo exposure to cartap induces tissue damage probably via oxidative stress in important vital organs of mice.
Assuntos
Antioxidantes/metabolismo , Inseticidas/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Tiocarbamatos/toxicidade , Administração Oral , Animais , Biomarcadores/sangue , Encéfalo/efeitos dos fármacos , Encéfalo/patologia , Relação Dose-Resposta a Droga , Inseticidas/administração & dosagem , Rim/efeitos dos fármacos , Rim/patologia , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Camundongos , Tiocarbamatos/administração & dosagemRESUMO
Dermatophytosis is a disease of global significance caused by pathogenic keratinolytic fungi called dermatophytes in both animals and humans. The recent taxonomy of dermatophytes classifies them into six pathogenic genera, namely Microsporum, Trichophyton, Epidermophyton, Nannizzia, Lophophyton and Arthroderma. It is because of the delayed diagnostic nature and low accuracy of dermatophyte detection by conventional methods that paved the path for the evolution of molecular diagnostic techniques, which provide the accurate and rapid diagnosis of dermatophytosis for an appropriate, timely antifungal therapy that prevents the nonspecific over-the-counter self-medication. This review focuses on the importance of rapid and accurate diagnosis of dermatophytosis, limitations of conventional methods, selection of targets in diagnosis, and factors affecting sensitivity and specificity of various molecular diagnostic technologies in the diagnosis of dermatophytosis. Generally, all the molecular techniques have a significant edge over the conventional methods of culture and microscopy in the dermatophytosis diagnosis. However, in mycology laboratory, the suitability of any molecular diagnostic technique in the diagnosis of dermatophytosis is driven by the requirement of time, economy, complexity, the range of species spectrum detected and the scale of diagnostic output required. Thus, various choices involved in the pursuit of a diagnosis of dermatophytosis are determined by the available conditions and the facilities in the laboratory.
Assuntos
Arthrodermataceae/patogenicidade , Técnicas de Diagnóstico Molecular/métodos , Tinha/diagnóstico , Animais , Antifúngicos/uso terapêutico , Arthrodermataceae/classificação , Humanos , Sensibilidade e Especificidade , Tinha/tratamento farmacológicoRESUMO
OBJECTIVE: This study aimed to assess the chondroprotective potential of atorvastatin in rat's cartilage explant culture model of osteoarthritis, stimulated by interleukin-1ß (IL-1ß). MATERIALS AND METHODS: The cartilage explants were treated with 20 ng/ml IL-1ß alone or with 20 ng/ml IL-1ß + various concentration of atorvastatin (1, 3, or 10 µM dissolved in DMSO) and incubated at 37 °C for 24 h. Also, control (0.25% DMSO), stimulated (20 ng IL-1ß) and treatment (atorvastatin 10 µM) cartilage explants were incubated without and with 1400W (10 µM). After 24 h of incubation, TNF-α, PGE2, MMP-13, TIMP-1, NO, and superoxide anion formation (O2(-)) concomitant with glycosaminoglycans (GAGs) were estimated in the medium. RESULTS: Atorvastatin inhibited IL-1ß-induced GAGs release, TNF-α, MMP-13, and O2(-) with no effect on TIMP-1 and NO. In addition, the source of NO in normal and atorvastatin-treated cartilage was eNOS, while for IL-1ß-stimulated cartilage it was iNOS. The cartilage degradation was associated with the combined effects of increased NO and O2 (-) rather than only NO. CONCLUSION: The present study suggests that atorvastatin has the ability to protect cartilage degradation following IL-1ß-stimulated cartilage in in vitro OA model and supports additional therapeutic application of atorvastatin in OA.
Assuntos
Anti-Inflamatórios/farmacologia , Atorvastatina/farmacologia , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Osteoartrite/metabolismo , Substâncias Protetoras/farmacologia , Animais , Cartilagem Articular/patologia , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Glicosaminoglicanos/metabolismo , Técnicas In Vitro , Interleucina-1beta/efeitos adversos , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Óxido Nítrico/metabolismo , Osteoartrite/induzido quimicamente , Osteoartrite/patologia , Ratos , Ratos Wistar , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: Sepsis commonly progresses to acute lung injury and is associated with high morbidity and mortality. Septic acute lung injury is characterized by severe oxidative stress response, remained refractory to present therapies, and new therapies need to be developed to improve further clinical outcomes. We determined the effect of betulinic acid (BA) on oxidative lung injury in mice using cecal ligation and puncture (CLP) model. MATERIALS AND METHODS: Five groups of mice (six in each group) received three pretreatments at 24-h interval before surgery. Surgery was done 1 h after last dosing. Sham and CLP control group mice received vehicle. BA was administered to other three groups of mice at 3, 10, and 30 mg/kg dose. Lung and plasma samples were collected for analysis by sacrificing the mice at 18 h of surgery. RESULTS: Compared with sham, CLP significantly increased total protein, nitrite, malondialdehyde, isoprostane, superoxide, protein carbonyl, oxidative stress index, inducible nitric oxide synthase protein, and histopathologic changes and reduced the superoxide dismutase, catalase activity, and total thiol levels in lungs and plasma, which were restored by BA pretreatment. CONCLUSIONS: BA pretreatment decreased the levels of oxidants, increased the levels of antioxidants in lungs and plasma thereby reducing the oxidative lung injury in CLP mice. Additionally, BA was found to scavenge the superoxide and nitric oxide radical in vitro. Thus, BA is suggested to be effective in treatment of oxidative lung injury in sepsis.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Lesão Pulmonar/prevenção & controle , Estresse Oxidativo/efeitos dos fármacos , Sepse/complicações , Triterpenos/uso terapêutico , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/metabolismo , Dinoprosta/análogos & derivados , Dinoprosta/metabolismo , Avaliação Pré-Clínica de Medicamentos , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Lesão Pulmonar/etiologia , Masculino , Malondialdeído/metabolismo , Camundongos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Nitritos/metabolismo , Triterpenos Pentacíclicos , Carbonilação Proteica/efeitos dos fármacos , Superóxidos/metabolismo , Triterpenos/farmacologia , Ácido BetulínicoRESUMO
BACKGROUND & OBJECTIVES: Osteoarthritis (OA) is a degenerative disease characterized by joint pain and progressive loss of articular cartilage. Entada pursaetha has been traditionally used in the treatment of inflammatory disease, liver ailment, etc. In this study we investigated suppressive effect of ethanolic extract of E. pursaetha (EPE) on monosodium iodoacetate (MIA)-induced osteoarthritis pain and disease progression by histopathological changes in joints in a rat model. METHODS: OA was induced in right knee of rat by intra-articular injection of 3 mg of MIA and characterized by pathological progression of disease and pain of affected joint. Spontaneous movements, mechanical, thermal and cold sensitivity were monitored at days 0 (before drug and MIA injection), 7, 14 and 21 of MIA administration. EPE (30, 100 and 300 mg/kg), vehicle or etoricoxib (10 mg/ kg; reference drug) were administered daily for 21 days by oral route. RESULTS: EPE at various doses significantly reduced mechanical, heat, cold hyperalgesia and increased the horizontal and vertical movements in intra-articular MIA injected rats. EPE prevented the damage to cartilage structure and reduced the cellular abnormalities. Articular cartilage of rats treated with EPE at 300 mg/kg group was almost normal with well-developed smooth surface and chondrocytes were distributed individually or arranged in column. INTERPRETATION & CONCLUSIONS: The present findings showed that the EPE was not only able to mitigate pain and hyperalgesia but also inhibited MIA-induced cartilage degeneration in vivo. EPE may have the potential to become therapeutic modality in the treatment of osteoarthritis. However, further studies need to be done to confirm these findings in other models and clinical trials.
Assuntos
Artrite Experimental/tratamento farmacológico , Osteoartrite/tratamento farmacológico , Dor/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Animais , Artrite Experimental/induzido quimicamente , Artrite Experimental/patologia , Cartilagem Articular/efeitos dos fármacos , Condrócitos/efeitos dos fármacos , Modelos Animais de Doenças , Fabaceae/química , Humanos , Injeções Intra-Articulares , Iodoacetatos/toxicidade , Masculino , Osteoartrite/induzido quimicamente , Osteoartrite/patologia , Dor/patologia , Extratos Vegetais/química , RatosRESUMO
The aim of the present study was to evaluate in vivo modulatory effect of S-methylisothiourea (SMT), a preferential inhibitor of inducible nitric oxide synthase (iNOS) on pain and pathology in the surgical model of osteoarthritis (OA) in rats. The OA was produced by the anterior cruciate ligament transection (ACLT) and medial meniscectomy (MMx) of right knee. SMT was administered 1 day prior to the production of OA and continued up to day 42 postoperation. Mechanical hyperalgesia, thermal hyperalgesia, tail flick latency after repeated flexion and extension of OA knee and knee diameter of right knee were determined at weekly intervals. Serum levels of IL-1ß, TNF-α and nitrite concentration were determined at the end of the experiment. Glycosaminoglycan (GAG) content, collagen content and histopathological evaluation of articular cartilage were also determined at the end of the experiment. SMT reduced mechanical hyperalgesia and the serum levels of IL-1ß, TNF-α and nitrite. Further, SMT reduced the loss of GAG from articular cartilage. Microscopically, SMT reduced the severity of the cartilage lesion. The results indicate the effectiveness of SMT in attenuating the pain and pathology of experimental OA phase by reducing the production of nitric oxide and interleukin-1ß and tumor necrosis factor-α, which are known to play a major role in the pathophysiology of OA.
Assuntos
Artralgia/tratamento farmacológico , Artralgia/patologia , Isotiurônio/análogos & derivados , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Osteoartrite/complicações , Análise de Variância , Animais , Ligamento Cruzado Anterior/cirurgia , Artralgia/etiologia , Técnicas Histológicas , Hiperalgesia/tratamento farmacológico , Interleucina-1beta/sangue , Isotiurônio/farmacologia , Masculino , Meniscos Tibiais/cirurgia , Nitritos/sangue , Ratos , Ratos Wistar , Fator de Necrose Tumoral alfa/sangueRESUMO
The purpose of this study was to determine the receptor subtype and the underlying mechanisms involved in the relaxant effect to leptin in mid- and late-pregnant mouse uterus. We determined the relative mRNA expression of receptor subtypes, eNOS, and BKCa channel by quantitative PCR and also the overall receptor expression by immunohistochemistry. Isometric tension studies were conducted to evaluate the effects of leptin and to delineate its mechanisms. A selective siRNA for the ObRb receptor was used to determine the participation of the receptor subtype in biochemical and molecular effects of leptin. The relaxant response to leptin was greater in mid-pregnancy compared to late pregnancy and was mediated by the activation of BKCa channels by eNOS-derived nitric oxide in an ObRb receptor-dependent manner. In comparison to mid-pregnancy, expression of short forms (mainly ObRa receptor) of the receptor was significantly increased in late pregnancy, whereas ObRb receptor expression was similar in both phases. The results of the study suggest that ObRb receptor mediates leptin-induced increase in eNOS expression and NO synthesis. Leptin-induced eNOS expression and activation cause cGMP-independent stimulation of BKCa channels causing uterine relaxation. Increased short forms of the receptors and reduced BKCa channels exert a negative effect on uterine relaxation in late pregnancy. Leptin may have a physiological role in maintaining uterine quiescence in mid-pregnancy and its reduced relaxant response in late gestation may facilitate labor. Further, ObRb receptor agonists may be useful in the management of preterm labor.
Assuntos
Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta , Leptina , Óxido Nítrico Sintase Tipo III , Óxido Nítrico , Receptores para Leptina , Transdução de Sinais , Útero , Animais , Feminino , Camundongos , Gravidez , Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta/metabolismo , Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta/genética , Leptina/farmacologia , Leptina/metabolismo , Relaxamento Muscular/efeitos dos fármacos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Receptores para Leptina/metabolismo , Receptores para Leptina/genética , Transdução de Sinais/efeitos dos fármacos , Útero/metabolismo , Útero/efeitos dos fármacosRESUMO
Ethion is a class II moderately toxic organothiophosphate pesticide. The main objective of this study was to evaluate the maternal and foetal toxicity of ethion in rats. Pregnant rats were divided into 5 groups. Group I served as control. Group II, III, IV, and V were orally administered with 0.86, 1.71, 3.43, and 6.9â¯mg/kg of ethion respectively, from gestational day (GD) 6-19. Dams were sacrificed on GD 20. Maternal toxicity was assessed by body weight gain, foetal resorptions, oxidative stress, liver and kidney function tests, and histopathology. Foetal toxicity was assessed by physical status, gross, teratological and histopathological examination. Ethion caused dose-dependent reduction in maternal body weight gain, increased resorptions, and reduced gravid uterine weights. Elevated MDA levels and altered levels of GSH, SOD and catalase were recorded in pregnant dam serum and tissues. SGOT, SGPT, total bilirubin, urea, uric acid, and creatinine were elevated in ethion groups indicating liver and kidney toxicity. Histology of uterus revealed myometrial degeneration and mucosal gland atrophy in uterus of pregnant dams and degenerative changes in placenta. It showed histological alterations in liver, kidney, and lungs. There was reduction in the foetal body weights and placental weights, and degenerative changes in the foetal liver and kidney. Gross evaluation of foetuses showed subcutaneous hematoma. Skeletal evaluation showed partial ossification of skull bones, costal separation, and agenesis of tail vertebrae, sternebrae, metacarpals and metatarsals. The findings reveal that prenatal exposure to ethion caused maternal and foetal toxicity in rats.
Assuntos
Rim , Fígado , Animais , Feminino , Gravidez , Ratos , Rim/efeitos dos fármacos , Rim/patologia , Fígado/efeitos dos fármacos , Fígado/patologia , Útero/efeitos dos fármacos , Útero/patologia , Estresse Oxidativo/efeitos dos fármacos , Etilenotioureia/toxicidade , Exposição Materna , Feto/efeitos dos fármacos , Feto/patologia , Tamanho do Órgão/efeitos dos fármacos , Ratos Wistar , Inseticidas/toxicidade , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Placenta/efeitos dos fármacos , Placenta/patologia , Reabsorção do Feto/induzido quimicamente , Troca Materno-Fetal , Desenvolvimento Fetal/efeitos dos fármacosRESUMO
Icariin is a flavonoid from plant belonging to the genus Epimedium, commonly known as Horny goat weed or Yin Yang Huo. The compound possesses multiple biological activities which are associated with the modulation of many signalling pathways, like NF-κB, Erk-p38-JNK, and release of various cytokines and growth factors. The present study determined wound healing potential of icariin in male Wistar rats. Icariin ointment (0%, 0.004%, 0.02%, 0.1% and 0.5%), was applied daily (b.i.d.) for 14 days onâ¯≈â¯400â¯mm2 cutaneous wound in different groups of rats. On day 14 post-wounding, 0.1% and 0.5% icariin treatment significantly (Pâ¯<â¯0.01 and Pâ¯<â¯0.001, respectively) increased wound contraction, as compared to control. Western blots revealed upregulation of IL-10 and downregulation of NF-κB and TNF-α. Increased expression of CD-31 showed abundance of microvessels in healing tissues after treatment with icariin. The MMP-2 and MMP-9 activities were reduced in icariin treated groups. Masson's trichrome staining revealed relatively better completion of re-epithelisation as well as increased deposition of well organised collagen fibres in the healing tissues compared to control. It is concluded that icariin has potential to accelerate cutaneous wound healing in rats.
Assuntos
Flavonoides/farmacologia , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Neovascularização Fisiológica/efeitos dos fármacos , Pele/irrigação sanguínea , Pele/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Animais , Regulação para Baixo/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Interleucina-10/metabolismo , Masculino , NF-kappa B/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Pele/citologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Ursolic acid (UA), an ursane-type pentacyclic triterpenoid commonly found in apple peels and holy basil has been shown to possess many beneficial effects. Renal fibrosis is a complication of kidney injury and associated with increased risk of morbidity and mortality. In our previous investigation, a lupane-type pentacyclic triterpenoid, betulinic acid (BA) was found to have protective effect on chronic kidney disease (CKD) and renal fibrosis. This prompted us to explore the therapeutic value of UA, a chemically related compound to BA in CKD. CKD was induced by feeding adenine with the feed at a concentration of 0.75% for 28 days. UA at the dose rate of 30â¯mg/kg in 0.5% carboxy methyl cellulose (CMC) was administered by oral route, simultaneously with adenine feeding for 28 days. Adenine feeding increased the kidney weight to body weight index, decreased the kidney function due to injury as indicated by increased markers like serum urea, uric acid, creatinine, cystatin C and neutrophil gelatinase-associated lipocalin (NGAL) and initiated the fibrotic response in kidney by increasing the profibrotic proteins viz. transforming growth factor-beta (TGF-ß), connective tissue growth factor (CTGF), fibronectin and collagen. However, treatment with UA reversed the damage induced by adenine as shown by reduced kidney injury and fibrosis markers which was further clearly evident in histological picture indicating the suitability of UA for use in CKD.
Assuntos
Rim/patologia , Insuficiência Renal Crônica/induzido quimicamente , Insuficiência Renal Crônica/tratamento farmacológico , Triterpenos/uso terapêutico , Adenina , Animais , Nitrogênio da Ureia Sanguínea , Colágeno Tipo I/metabolismo , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Creatinina/sangue , Cistatina C/metabolismo , Fibronectinas/metabolismo , Fibrose , Hidroxiprolina/metabolismo , Rim/efeitos dos fármacos , Lipocalina-2/sangue , Masculino , Tamanho do Órgão/efeitos dos fármacos , Ratos Wistar , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/patologia , Fator de Crescimento Transformador beta1/metabolismo , Triterpenos/química , Triterpenos/farmacologia , Ácido Úrico/sangue , Ácido UrsólicoRESUMO
BACKGROUND: Most chronic kidney diseases (CKDs), regardless of the nature of the initial injury, progress to end-stage renal disease (ESRD) characterized by fibrosis with irreversible loss of tissue and function. Thus, improved and more effective therapies are critical. Betulinic acid (BA), a pentacyclic triterpene is a compound in the pipeline of anti-cancer drug development. It has been shown to a possess variety of beneficial effects in many disease conditions. However, its efficacy against CKD is yet to be explored. OBJECTIVE: The present study was undertaken to investigate the effect of BA on renal fibrosis in the rat model of adenine-induced CKD. RESULTS: CKD rats gained significantly less weight during the experimental period when compared to control rats and BA treatment did not significantly increase the weight gain in CKD rats. CKD rats showed elevated levels of serum blood urea nitrogen (BUN), creatinine and uric acid along with increased levels of kidney injury markers such as cystatin C and neutrophil gelatinase-associated lipocalin (NGAL). Further, in comparison to control rats, kidney samples from CKD rats revealed increased profibrotic protein levels like transforming growth factor-beta (TGF-ß), connective tissue growth factor (CTGF), fibronectin, collagen type I and hydroxyproline indicating a progressive fibrotic response. These data are further fortified by histological findings where kidney damage and fibrosis are clearly evident as dilatation of tubules, glomerular degeneration and vacuolation along with deposition of collagen fibers. However, the above-mentioned findings in CKD rats were significantly reversed by BA-treatment revealing its nephroprotective potential and anti-fibrotic activity. CONCLUSION: The biochemical mechanism of the nephroprotective and anti-fibrotic effect of BA in the adenine-induced CKD rats might be mediated by inhibition of pro-fibrotic protein production thereby hindering the kidney tissue damage along with improvement in kidney function. Thus, BA could be an adjunct agent to retard fibrosis in CKD subjects.
Assuntos
Fibrose/tratamento farmacológico , Insuficiência Renal Crônica/tratamento farmacológico , Triterpenos/farmacologia , Animais , Peso Corporal/efeitos dos fármacos , Fibrose/prevenção & controle , Regulação da Expressão Gênica/efeitos dos fármacos , Rim/efeitos dos fármacos , Rim/patologia , Estrutura Molecular , Tamanho do Órgão/efeitos dos fármacos , Triterpenos Pentacíclicos , Ratos , Triterpenos/química , Ácido BetulínicoRESUMO
Ursolic acid (UA; 3b-hydroxy-12-urs-12-en-28-oic acid), a natural pentacyclic triterpenoid carboxylic acid, has been known to possess potent anti-inflammatory, antioxidant, and antinociceptive effects in various animal models. Therefore, this study was designed to investigate the antihyperalgesic, anti-inflammatory, and antioxidant effects of UA at 5, 10, and 20 mg/kg of doses via per os (p.o.) route for 14 days in chronic constriction injury (CCI)-induced neuropathic pain in rats. Pain behavior in rats was evaluated before and after UA administration via mechanical and heat hyperalgesia. CCI caused significant increase in levels of pro-inflammatory cytokines and oxido-nitrosative stress. In addition, significant increase in myeloperoxidase, malondialdehyde, protein carbonyl, nitric oxide (NO), and total oxidant status (TOS) levels in sciatic nerve and spinal cord concomitant with mechanical and heat hyperalgesia is also noted for CCI-induced neuropathic pain. Administration of UA significantly reduced the increased levels of pro-inflammatory cytokines and TOS. Further, reduced glutathione is also restored by UA. UA also showed in vitro NO and superoxide radical scavenging activity. UA has a potential in attenuating neuropathic pain behavior in CCI model which may possibly be attributed to its anti-inflammatory and antioxidant properties.
Assuntos
Neuralgia/tratamento farmacológico , Triterpenos/farmacologia , Animais , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Constrição , Citocinas/metabolismo , Modelos Animais de Doenças , Hiperalgesia/tratamento farmacológico , Hiperalgesia/metabolismo , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Masculino , Malondialdeído/metabolismo , Neuralgia/metabolismo , Óxido Nítrico/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Peroxidase/metabolismo , Carbonilação Proteica/efeitos dos fármacos , Ratos , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/metabolismo , Medula Espinal/efeitos dos fármacos , Medula Espinal/metabolismo , Superóxidos/metabolismo , Ácido UrsólicoRESUMO
Diarylheptanoids, a group of plant secondary metabolites are increasingly recognized as potential therapeutic agents. The aim of study was to ascertain the anti-inflammatory profile of diarylheptanoids from Alnus nepalensis against lipopolysaccharide (LPS)-induced inflammation in macrophages and endotoxic shock in mice. Extracts prepared from dried leaves of A. nepalensis using standard solvents were tested against LPS-induced inflammation in macrophages. Among all, butanol extract (ANB) has shown most significant inhibition of pro-inflammatory cytokines without any cytotoxicity. HPLC analysis of ANB showed the presence of diarylheptanoids. The diarylheptanoids were further isolated and tested in-vitro for anti-inflammatory activity. Treatment of isolated diarylheptanoids (HOG, ORE and PLS) was able to reduce the production and mRNA level of pro-inflammatory cytokines (TNF-α and IL-6). Furthermore, we demonstrated that it inhibited the expression of NF-kB protein in LPS-induced inflammation in macrophages. In-vivo efficacy and safety profile of ANB revealed that oral treatment of ANB was able to improve the survival rate, and inhibited the production of pro-inflammatory cytokines in serum, attenuated vital organ injury in a dose dependent manner without any toxic effect at higher dose in mice. The results suggest that diarylheptanoids from A. nepalensis can be considered as potential therapeutic candidates for the management of inflammation related diseases.
Assuntos
Anti-Inflamatórios/administração & dosagem , Diarileptanoides/administração & dosagem , Inflamação/prevenção & controle , Macrófagos Peritoneais/efeitos dos fármacos , Choque Séptico/tratamento farmacológico , Alnus/imunologia , Animais , Linhagem Celular , Diarileptanoides/isolamento & purificação , Inflamação/induzido quimicamente , Interleucina-6/genética , Interleucina-6/metabolismo , Lipopolissacarídeos/imunologia , Macrófagos Peritoneais/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Extratos Vegetais , Folhas de Planta , Choque Séptico/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Deferoxamine has shown cutaneous wound healing potential by increased neovascularization. We hypothesized that topically applied deferoxamine facilitates wound healing in diabetic rats by modulating important cytokines and growth factors that take part in healing processes in a time-dependent manner. Diabetes was induced in male Wistar rats by streptozotocin and wound was created under pentobarbitone anesthesia. The diabetic rats were divided into two groups, of which one (control) was treated with ointment base and other with deferoxamine ointment (0.1%). Wound closure measurement and tissue collection were done on days 3, 7, 14 and 19 post-wounding. The relative expressions of hypoxia-inducible factor 1-alpha (HIF-1α), vascular endothelial growth factor (VEGF), stromal cell-derived factor 1-alpha (SDF-1α), transforming growth factor beta 1 (TGF-ß1), tumor necrosis factor-alpha (TNF-α), matrix metalloproteinase-9 (MMP-9), interleukin-1 beta (IL-1ß) and interleukin-10 (IL-10) mRNA and proteins were determined in the wound tissues. CD-31 staining and collagen content were evaluated by immunohistochemistry and picrosirius red staining, respectively. Histological changes were assessed by H&E staining. The per cent wound closure was significantly higher from day 7 onwards in deferoxamine-treated rats. HIF-1α, VEGF, SDF-1α, TGF-ß1, IL-10 mRNA and their protein levels were significantly higher on days 3, 7 and 14 in deferoxamine-treated rats. The mRNA expression and protein levels of TNF-α, MMP-9 and IL-1ß were progressively and markedly reduced in deferoxamine-treated rats. The collagen deposition and formation of blood vessels were greater in deferoxamine-treated rats. It is suggested that topical application of deferoxamine ointment might be useful in cutaneous wound healing in diabetic patients.
Assuntos
Citocinas/metabolismo , Desferroxamina/farmacologia , Diabetes Mellitus Experimental/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Pele/efeitos dos fármacos , Pele/fisiopatologia , Cicatrização/efeitos dos fármacos , Animais , Citocinas/genética , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Experimental/fisiopatologia , Regulação da Expressão Gênica/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos WistarRESUMO
BACKGROUND: Oxidative stress is one of the main causes of pain and cartilage degradation in osteoarthritis. This study on atorvastatin, a HMG-CoA reductase inhibitor used in the treatment of hypercholesterolemia and prevention of coronary heart disease aimed to investigate its effect on hyperalgesia and cartilage damage in monosodium iodoacetate (MIA)-induced osteoarthritis model in rats. METHODS: Osteoarthritis was induced by a single intra-articular injection of 3mg MIA. After daily administration of atorvastatin (3, 10 and 30 mg/kg) for 20 days by oral gavage, pain was assessed on days 0, 1, 3, 7, 14 and 21. Histopathology of ipsilateral knee joint; oxidative markers and antioxidants in plasma were assessed on day 21. RESULTS: Atorvastatin attenuated hyperalgesia. The increased level of lipid peroxidation, superoxide, protein carbonyl; decreased activity of catalase, glutathione-S-transferase, reduced glutathione and total thiol levels in MIA rats were restored to the normal levels, however, superoxide dismutase and nitric oxide levels remained unaltered by atorvastatin. Further, atorvastatin reduced the MIA-induced histopathological alteration in the knee joint. CONCLUSION: Our study demonstrated that atorvastatin attenuates MIA-induced osteoarthritic pain and protect cartilage degradation through inhibition of oxidative stress suggesting its importance in osteoarthritic pain management.
Assuntos
Atorvastatina/uso terapêutico , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Ácido Iodoacético/toxicidade , Osteoartrite/tratamento farmacológico , Dor/tratamento farmacológico , Animais , Atorvastatina/farmacologia , Relação Dose-Resposta a Droga , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Injeções Intra-Articulares , Masculino , Osteoartrite/induzido quimicamente , Osteoartrite/patologia , Dor/induzido quimicamente , Dor/patologia , Medição da Dor/efeitos dos fármacos , Medição da Dor/métodos , Ratos , Ratos Wistar , Resultado do TratamentoRESUMO
Atorvastatin is an HMG-CoA reductase inhibitor used in the treatment of hypercholesterolemia and prevention of coronary heart disease. Oxidative stress is considered to be one of the main causes of neuropathic pain after nerve injury. This study aimed to investigate the effect of atorvastatin on oxidative stress and hyperalgesia in chronic constriction injury (CCI) model of neuropathic pain. Pain behaviour in rats was evaluated before and after atorvastatin administration using mechanical and heat hyperalgesia. The markers for oxidative stress in sciatic nerve, spinal cord and pre-frontal cortex (PFC) area of brain were biochemically detected in vehicle and atorvastatin-treated neuropathic CCI rats. Atorvastatin attenuated hyperalgesia. We found a significant increase in malondialdehyde (MDA), nitric oxide (NO), superoxide anion (O2(-)) and protein carbonyl along with a reduction in catalase (CAT), reduced glutathione (GSH), total thiol (SH) and glutathione-S-transferase (GST) and; increase in superoxide dismutase (SOD) levels in the sciatic nerve, spinal cord and PFC of the CCI-induced neuropathic rats. Reduced levels of enzymatic and non enzymatic antioxidants were restored by atorvastatin. The levels of MDA, O2(-), and protein carbonyl in these tissues were significantly reduced in the atorvastatin-treated CCI rats compared to the untreated CCI rats. Our study demonstrated that atorvastatin attenuates neuropathic pain through inhibition of oxidative stress in sciatic nerve, spinal cord and brain suggesting antioxidants as potential drugs in neuropathic pain management. This study provides a new application of atorvastatin in treatment of neuropathic pain.
Assuntos
Ácidos Heptanoicos/farmacologia , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Neuralgia/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Pirróis/farmacologia , Nervo Isquiático/efeitos dos fármacos , Medula Espinal/efeitos dos fármacos , Animais , Antioxidantes/uso terapêutico , Atorvastatina , Regulação para Baixo , Hiperalgesia/complicações , Hiperalgesia/tratamento farmacológico , Masculino , Ratos WistarRESUMO
Oxidative injury is markedly responsible for wound complications in diabetes mellitus. The biological actions of bilirubin may be relevant to prevent oxidant-mediated cell death, as bilirubin application at a low concentration scavenges reactive oxygen species. Hence, we hypothesized that topical bilirubin application might improve wound healing in diabetic rats. Diabetes was induced in adult male Wistar rats, which were divided into two groups, i.e., diabetic control and diabetic treated. Non-diabetic healthy rats were also taken as healthy control group. Wound area was measured on days 3, 7, 14, and 19 post-wounding. The levels of malondialdehyde (MDA) and reduced glutathione (GSH) and the activities of glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) were estimated in the granulation tissue. There was a significant increase in percent wound closure in healthy control and diabetic treated rats on days 7, 14, and 19, as compared to diabetic control rats on days 7, 14, and 19. There was significant decrease in MDA levels on days 7, 14, and 19 in diabetic treated rats, as compared to diabetic control rats. Levels of GSH were significantly increased on days 3, 7, 14, and 19 in diabetic treated rats, as compared to diabetic control rats. GPx, SOD, and CAT activities were significantly higher on days 3, 7, and 14 in diabetic treated rats, as compared to diabetic control rats. The findings indicate that bilirubin is effective in reducing the oxidant status in wounds of diabetic rats which might have accelerated wound healing in these rats.
Assuntos
Antioxidantes/administração & dosagem , Bilirrubina/administração & dosagem , Diabetes Mellitus Experimental/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Animais , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Masculino , Estresse Oxidativo/fisiologia , Ratos , Ratos Wistar , Estreptozocina , Cicatrização/fisiologiaRESUMO
Atorvastatin is a 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase inhibitor used in treatment of hypercholesterolemia and prevention of coronary heart disease. The aim of this study is to investigate the antihyperalgesic and anti-inflammatory effects of atorvastatin (3, 10, and 30 mg/kg by oral gavages for 14 days) in chronic constriction injury (CCI) model of neuropathic pain in rats. CCI caused significant increase in tumor necrosis factor-α, interleukin 1 beta, prostaglandin E2, along with matrix metalloproteases (MMP-2) and nerve growth factor (NGF) levels in sciatic nerve and spinal cord concomitant with mechanical and thermal hyperalgesia, which were significantly reduced by oral administration of atorvastatin for 14 days as compared to CCI rats. Our study demonstrated that atorvastatin attenuates neuropathic pain through inhibition of cytokines, MMP-2, and NGF in sciatic nerve and spinal cord suggesting that atorvastatin could be an additional therapeutic strategy in management of neuropathic pain.
Assuntos
Ácidos Heptanoicos/uso terapêutico , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Neuralgia/tratamento farmacológico , Pirróis/uso terapêutico , Nervo Isquiático/metabolismo , Medula Espinal/metabolismo , Analgesia , Animais , Anti-Inflamatórios/uso terapêutico , Atorvastatina , Constrição , Dinoprostona/biossíntese , Modelos Animais de Doenças , Interleucina-1beta/biossíntese , Masculino , Metaloproteinase 2 da Matriz/biossíntese , Fator de Crescimento Neural/biossíntese , Medição da Dor , Ratos , Ratos Wistar , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Much information is available on the role of nitric oxide (NO) in osteoarthritis (OA). However, its role has not been studied in the monosodium iodoacetate (MIA)-induced model of osteoarthritic pain. The present study was undertaken in rats to investigate the effect of iNOS inhibitor S-methylisothiourea (SMT) in MIA-induced osteoathritic pain and disease progression in rats. Osteoarthritis was produced by single intra-articular injection of the MIA in the right knee joint on day 0. Treatment groups were orally gavazed with different doses of SMT (10, 30 and 100mg/kg) and etoricoxib (10mg/kg) daily for 21 days. On days 0, 3, 7, 14 and 21, pain was measured and histopathology of right knee joint was done on day 21. SMT produced analgesia in a dose-dependent manner as shown by mechanical, heat hyperalgesia, knee vocalization, knee squeeze test, and spontaneous motor activity test. SMT reduced NO production in synovial fluid. Histopathological findings indicated that SMT reduced disease progression as evident from complete cartilage formation in rats treated with SMT at 30 mg/kg. In conclusion, the results indicate that SMT attenuates the MIA-induced pain and histopathological changes in the knee joint. The antinociceptive and antiarthritic effects of SMT were mediated by inhibiting cartilage damage and suppression of NO in synovial fluid. It is suggested that SMT has potential as a therapeutic modality in the treatment of osteoarthritis.