RESUMEN
Wounds treated with TiO2 nanoparticles (TiO2-NPs) show an improvement in healing time. However, little is known about the parameters that can contribute to this result. On the other hand, the treatment of wounds with polyphenols is widely known. These compounds are found in the peel of Annona crassiflora fruit and have antioxidant, analgesic and anti-inflammatory properties. In this study, we evaluated the healing effect of TiO2 nanocrystals (TiO2-NCs), polyphenolic fractions obtained from ethanolic extract of A. crassiflora fruit peel (PFAC) and mix (PFAC + TiO2-NCs) on the parameters of wound closure, inflammation, collagen deposition, metalloproteinase activity (MMPs) and angiogenesis. TiO2-NCs and PFAC have activity for wound healing, showed anti-inflammatory action and a shorter wound closure time. These treatments also contributed to increased collagen deposition, while only treatment with TiO2-NCs increased MMP-2 activity, parameters essential for the migration of keratinocytes and for complete restoration of the injured tissue. The combination of PFAC + TiO2-NCs reduced the effectiveness of individual treatments by intensifying the inflammatory process, in addition to delaying wound closure. We conclude that the interaction between the hydroxyl groups of PFAC polyphenols with TiO2-NCs may have contributed to difference in the healing activity of skin wounds.
Asunto(s)
Annona , Nanopartículas , Annona/química , Antiinflamatorios/farmacología , Colágeno , Nanopartículas/química , Polifenoles/farmacología , Titanio , Cicatrización de HeridasRESUMEN
Annona crassiflora Mart. is a species native to the Cerrado biome, whose fruit is known as araticum or marolo. Plant parts are widely used in folk medicine to treat inflammation and pain associated with rheumatism, wounds, venereal diseases, snakebites, and microbial infections. Thus, we investigated a fraction rich in phenolic compounds (PCAc) obtained from the crude extract of the peel of these fruits on non-cytotoxic, anti-inflammatory, antioxidant, and collagen biosynthesis properties in the healing of wounds induced on the back of BALB/c mice. For the control group, the induced wounds were not treated and for the others, wounds were treated topically with vehicle or vehicle plus PCAc. Both fractions contained in PCAc demonstrated effective protection on fibroblasts. We highlight the effect of the ethyl acetate fraction which, in addition to the protective effect, has a proliferative activity on these cells. In addition, PCAc caused improvement in healing after 7 days of treatment and in the longest period of treatment with PCAc (7, 14, and 21 days) there was a greater contraction of the wound, accompanied by resolution of the inflammatory process, antioxidant defense, increasing collagen synthesis, and modulation of metalloproteinases. PCAc demonstrated better re-epithelialization and organization of the dermis at the end of treatment. The changes promoted by the phenolic compounds of A. crassiflora were important in the healing process, especially in activities related to inflammation, oxidative stress, and fibrogenesis.
Asunto(s)
Annona , Animales , Antioxidantes/farmacología , Fibroblastos , Ratones , Extractos Vegetales/farmacología , Cicatrización de HeridasRESUMEN
α-zingiberene is a phytochemical of the sesquiterpenes class, the major constituent of the essential oil from the leaves of Casearia sylvestris, a plant widely used in traditional medicine for the treatment of inflammatory diseases, tumours, and bacterial infections. In the present study, we evaluated the effects of daily administration of α-zingiberene (0.01, 0.1 and 1 µg diluted in 10 µl of 0.5% DMSO) on the inflammatory, angiogenic, and fibrogenic components, induced by subcutaneous sponge implants in an animal model. Treatment with sesquiterpene resulted in a reduction in macrophage activation, as well as in mean blood vessels and in the activity of metalloproteinases 2 and 9. Furthermore, it resulted in an increase in collagen deposition near the implants. These results show the therapeutic potential of α-zingiberene in the treatment of pathologies, in which processes such as inflammation and angiogenesis are exacerbated, or even for the treatment of chronic wounds.
Asunto(s)
Casearia , Aceites Volátiles , Sesquiterpenos , Ratones , Animales , Aceites Volátiles/farmacología , Hojas de la Planta , Sesquiterpenos/farmacología , Colágeno , Neovascularización Patológica/tratamiento farmacológico , Extractos Vegetales/farmacologíaRESUMEN
Jararhagin-C (Jar-C) is a disintegrin-like protein, isolated from the venom of B. jararaca, with affinity for α2ß1 integrin and the ability to incite processes such as angiogenesis and collagen deposition in vivo. Thus, we raised the hypothesis that this protein could be used as a therapeutic strategy for stimulating the healing of excisional wounds in mice. Four wounds were made on the back of Swiss mice, treated with daily intradermal injections of PBS (control group) or Jar-C (200 ng). Ten animals from each experimental group were euthanized and the tissue from the wounds and skin around them were collected for further biochemical, histological and molecular analysis. Wounds treated with Jar-C showed a faster closure rate, accompanied by a reduction in neutrophil infiltrate (MPO), pro-inflammatory cytokine levels (TNF, CXCL1 and CCL2) and an accumulation of macrophages in the analyzed tissues. It was also observed a greater expression of genes associated with the phenotype of alternatively activated macrophages (M2). Concomitantly, the administration of Jar-C holds an angiogenic potential, increasing the density of blood vessels and the synthesis of pro-angiogenic cytokines (VEGF and FGF). We also observed an increase in collagen deposition, accompanied by higher levels of the pro-fibrogenic cytokine TGF-ß1. Our data suggests Jar-C stimulates wound healing through stimulation of M2-like macrophage, angiogenesis and collagen deposition. Jar-C may be explored as a therapeutic strategy for wound healing, including the treatment of chronic wounds, where processes such as inflammation, angiogenesis and the deposition / remodeling of the matrix constituents are unregulated.
Asunto(s)
Colágeno , Venenos de Crotálidos , Desintegrinas , Neovascularización Fisiológica , Cicatrización de Heridas , Animales , Humanos , Ratones , Supervivencia Celular/efectos de los fármacos , Colágeno/metabolismo , Venenos de Crotálidos/química , Citocinas/genética , Citocinas/metabolismo , Desintegrinas/química , Desintegrinas/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana , Leucocitos/efectos de los fármacos , Leucocitos/fisiología , Macrófagos , Neovascularización Fisiológica/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Cicatrización de Heridas/efectos de los fármacos , Veneno de Bothrops JararacaRESUMEN
BACKGROUND AND AIM: Maytenus ilicifolia has analgesic, healing, antioxidant and anti-inflammatory properties. This study evaluated effect of the hydroalcoholic extract of M. ilicifolia leaves on skin wound repair. EXPERIMENTAL PROCEDURE: Wounds were induced on mice and treated with the extract. The treatment was performed daily, until day 7 after wound induction. Wound closure was measured and the features of the repaired tissue were investigated, including mast cell quantification, neutrophil and macrophage activities, collagen deposition, angiogenesis, and pro-metalloproteases and metalloproteases 2 and 9 activity (pro-MMPs and MMPs). RESULTS AND CONCLUSION: The M. ilicifolia extract accelerated the closure of wounds. The extract at a concentration of 4% was found to be effective, presenting anti-inflammatory effects and hemoglobin increased, along with increased soluble, total and type III collagens in the wound. In addition, there was an increase in pro-MMP9 and MMP9 activity after day 7th of treatment. The phenolic compounds and tannins present in this plant could be associated with the anti-inflammatory and healing activities observed in this study. Therefore, the ability to modulate essential parameters for accelerated and adequate healing as shown here suggests that the use of standardised extracts of M. ilicifolia and its fractions enriched in polyphenols may represent a therapeutic strategy for the treatment of wounds.
RESUMEN
Drimys brasiliensis (Winteraceae) has been investigated in traditional medicine for its anti-inflammatory properties to treat gastric ulcers and allergic and respiratory system diseases as well as for cancer treatment. In this work, we investigate the ability of the sesquiterpene polygodial, isolated from D. brasiliensis stem barks, to modulate the chronic inflammatory response induced by polyester-polyurethane sponge implants in C57BL/6J mice. Daily treatment with polygodial inhibited the macrophage content in the implants as determined by the activity of the N-acetyl-ß-d-glucosaminidase enzyme as well as decreased the levels of CXCL1/KC and CCL2/JE/MCP-1 pro-inflammatory chemokines and the presence of mast cells along the formed fibrovascular tissue. Similarly, the deposition of a new extracellular matrix (total collagen and type I and III collagen fibers) as well as the production of the TGF-ß1 cytokine were attenuated in implants treated with polygodial, showing for the first time its antifibrogenic capacity. The hemoglobin content, the number of newly formed vessels, and the levels of VEGF cytokine, which were used as parameters for the assessment of the neovascularization of the implants, did not change after treatment with polygodial. The anti-inflammatory and antifibrogenic effects of polygodial over the components of the granulation tissue induced by the sponge implant indicate a therapeutic potential for the treatment of inflammatory diseases associated with the development of fibrovascular tissue.
Asunto(s)
Regulación hacia Abajo , Drimys/química , Inflamación/prevención & control , Sesquiterpenos/aislamiento & purificación , Winteraceae/química , Animales , Fibrosis/prevención & control , Ratones , Ratones Endogámicos C57BLRESUMEN
Trypanosoma cruzi P21 protein (P21) is a putative secreted and immunomodulatory molecule with potent bioactive properties such as induction of phagocytosis and actin cytoskeleton polymerization. Despite the bioactive properties described so far, the action of P21 on parasite replication in muscle cell lineage or T. cruzi parasitism during acute experimental infection is unclear. We observed that recombinant P21 (rP21) decreased the multiplication of T. cruzi in C2C12 myoblasts, phenomenon associated with greater actin polymerization and IFN-γ and IL-4 higher expression. During experimental infection, lower cardiac nests, inflammatory infiltrate and fibrosis were observed in mice infected and treated with rP21. These results were correlated with large expression of IFN-γ counterbalanced by high levels of IL-10, which was consistent with the lower cardiac tissue injury found in these mice. We have also observed that upon stress, such as that induced by the presence of the IFN-γ cytokine, T. cruzi produced more P21. The effect of P21 in controlling the replication of T. cruzi, may indicate an evolutionary mechanism of survival developed by the parasite. Thus, when subjected to different stress conditions, the protozoan produces more P21, which induces T. cruzi latency in the host organism, enabling the protozoan to evade the host's immune system.
Asunto(s)
Péptidos y Proteínas de Señalización Intercelular/metabolismo , Malaria/parasitología , Mioblastos/parasitología , Miocardio/patología , Proteínas Protozoarias/metabolismo , Trypanosoma cruzi/fisiología , Enfermedad Aguda , Animales , Línea Celular , Interacciones Huésped-Parásitos , Humanos , Evasión Inmune , Péptidos y Proteínas de Señalización Intercelular/genética , Interferón gamma/metabolismo , Malaria/inmunología , Ratones , Ratones Endogámicos C57BL , Modelos Animales , Carga de Parásitos , Proteínas Protozoarias/genéticaRESUMEN
Obesity is characterized by the excess of body fat and, therefore, may cause musculoskeletal alterations that can negatively influence the tendons. Such overweight-influenced alterations are exercise sensitive though. Morphological and biochemical alterations were reported in the calcaneal tendon of mice submitted to a lipid-rich diets along with practicing exercises, with the following groups: normal diet without exercise (ND), normal diet with exercise (NDex), lipid-rich diet without exercise (LD), lipid-rich diet without exercise (LDex). The calcaneal tendons were removed and subjected to histological and biochemical analysis. Layers of the tissue were stained with Hematoxylin and Eosin, Picrosirius Red and Von Kossa while a protein dosage was conduce by the Bradford method. The morphologicals analysis there was no statistical difference concerning the number of fibroblasts among the groups. Groups submitted to exercises showed higher amount of collagen and non-collagenous protein deposition. The lipid-rich diet without exercse group had a more disorganized collagen matrix with intense basophilia. The same group had areas of calcification confirmed by Von Kossa technique. Practicing physical activity, such as swimming, can improve the changes caused in the calcaneal tendon in mice submitted to a lipid-rich diets, having a better collagen organization and the synthesis.
Asunto(s)
Tendón Calcáneo/metabolismo , Colágeno/metabolismo , Grasas de la Dieta/metabolismo , Lípidos/administración & dosificación , Metaloproteinasa 2 de la Matriz/metabolismo , Condicionamiento Físico Animal , Natación , Animales , Grasas de la Dieta/administración & dosificación , Masculino , RatonesRESUMEN
Alternagin-C (ALT-C), a disintegrin-like protein obtained from the venom of Bothrops alternatus, is able to modulate cellular behaviors such as adhesion, migration and proliferation, as well as the production of various growth factors via α2ß1 integrin, important processes during inflammation, angiogenesis and fibrogenesis, which although appear as distinct events, act concomitantly in several chronic inflammatory diseases. Our objective was to investigate the effects of ALT-C on components of the sponge-induced inflammatory response in balb/c mice. The polyester-polyurethane sponges were implanted in mice's subcutaneous layer of the dorsal region and daily injected with saline (control group) or ALT-C (10, 100 or 1000â¯ng). Nine days after implantation the implants were removed and processed. ALT-C inhibited the inflammatory response, observed through mast cell reduction, NAG-activity and also by the inhibition of TNF-α, CXCL-1 and CCL2/JE/MCP-1 cytokines. ALT-C was also able to reduce hemoglobin content, number of vessels and the concentrations of VEGF and FGF cytokines. Finally, at its highest dose (1000â¯ng), ALT-C increased all evaluated markers associated with fibrogenesis (collagen production and TGF-ß1 levels). All these factors reveal that ALT-C is a strong candidate to be exploited in the development of anti-inflammatory and anti-angiogenic therapies in chronic inflammatory processes.
Asunto(s)
Bothrops/metabolismo , Colágeno/metabolismo , Venenos de Crotálidos/farmacología , Desintegrinas/farmacología , Inflamación/tratamiento farmacológico , Neovascularización Patológica/tratamiento farmacológico , Animales , Hemoglobinas/metabolismo , Inflamación/metabolismo , Masculino , Mastocitos/efectos de los fármacos , Mastocitos/metabolismo , Ratones , Ratones Endogámicos BALB C , Neovascularización Patológica/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
IL-9 is a pleiotropic cytokine, recently recognized as belonging to Th9 cells that are involved in various pathologies. We aimed to evaluate the role of IL-9 in the course of hepatic and renal fibrosis. Female C57BL/6 mice were treated subcutaneously with IL-9 10 ng/mouse and 20 ng/mouse for 40 days, alternating every 5 days each application, the negative control of which was treated with PBS and positive control with CCL4. IL-9 demonstrated fibrogenic activity, leading to increased collagen I and III deposition in both liver and kidney, as well as triggering lobular hepatitis. In addition, IL-9 induced an inflammatory response with recruitment of lymphocytes, neutrophils, and macrophages to both organs. The inflammation was present in the region of the portal and parenchymal zone in the liver and in the cortical and medullary zone in the kidney. IL-9 deregulated liver and kidney antioxidant activities. Our results showed that IL-9 was able to promote hepatorenal dysfunction. Moreover, IL-9 poses as a promising target for therapeutic interventions.
Asunto(s)
Fibrosis/etiología , Interleucina-9/efectos adversos , Riñón/patología , Hígado/patología , Animales , Colágeno/metabolismo , Femenino , Inflamación/inducido químicamente , Inflamación/patología , Riñón/fisiología , Hígado/fisiología , Ratones , Ratones Endogámicos C57BLRESUMEN
A polyphenol-enriched fraction from Annona crassiflora fruit peel (PEF-Ac) containing chlorogenic acid, epi-catechin, procyanidins B2 and C1, quercetin-glucoside, kaempferol, and caffeoyl-glucoside was investigated for its anti-inflammatory, pro-angiogenic, and profibrogenic potential in the healing of cutaneous wounds. Four wounds were performed on the back of C57 mice and the lesions were treated with the vehicle (Vaseline and lanolin) and PEF-Ac at concentrations of 2%, 4%, and 6% for 4 and 7 d. Neutrophils and macrophages activities were evaluated indirectly by the activity of myeloperoxidase and N-acetyl-ß-D-glycosaminidase, angiogenesis was evaluated by hemoglobin dosing and vessel count in histological sections, and collagen deposition was assessed from histological sections stained with picrosirius red. PEF-Ac demonstrated anti-inflammatory activity, with reduced activities of neutrophil and macrophage in the cutaneous wounds. In addition, there was an increase in the synthesis of types I and III collagen, as well as in the percentage of wound closure, mainly after 4 d of treatment. On the other hand, PEF-Ac did not present an effective pro-angiogenic activity. A. crassiflora fruit peel showed anti-inflammatory and profibrogenic properties, indicating a promising natural source of bioactive molecules for treatment of cutaneous wounds.
Asunto(s)
Annona/química , Antiinflamatorios no Esteroideos/farmacología , Extractos Vegetales/farmacología , Polifenoles/farmacocinética , Piel/lesiones , Cicatrización de Heridas/efectos de los fármacos , Acetilglucosaminidasa/metabolismo , Inductores de la Angiogénesis/farmacología , Animales , Colágeno/metabolismo , Hemoglobinas/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Infiltración Neutrófila/efectos de los fármacos , Peroxidasa/metabolismo , Piel/efectos de los fármacos , Piel/inmunología , Piel/metabolismoRESUMEN
Jararhagin, a metalloprotease from Bothrops jararaca snake venom, is a toxin containing the metalloproteinase, disintegrin-like and cysteine-rich domains; it causes acute inflammation and damage to vascular tissue. However, the actions of these domains on key components of chronic inflammation have not been determined. Our aim was to investigate the effects of jararhagin (Jar), jararhagin-C (Jar-C) and o-phenantrolin-treated jararhagin (Jar-Phe), on inflammatory response, blood vessel formation and extracellular matrix deposition in the murine sponge model. The polyether-polyurethane sponge matrix was implanted into Balb/c mice and injected daily with Jar (400â¯ng), Jar-Phe (400â¯ng), Jar-C (200â¯ng) or saline (control). Nine days after implantation, the sponge discs were removed and processed. In the Jar-treated implants, some of inflammatory markers (N-acetyl-ß-d-glucosaminidase activity, CCL2 and TNF-α) and TGF-ß1 levels were higher compared with the control group. In the Jar-C group, the inflammatory markers myeloperoxidase activity and CXCL1 were higher compared with the control. In this group, VEGF levels and collagen deposition were also higher. Jar-Phe treatment was able to inhibit the activity and/or production of MPO, CXCL1, CCL2 and TGF-ß. The differential effects of these proteins in modulating the main components of fibrovascular tissue may be exploited in the management fibroproliferative diseases.
Asunto(s)
Venenos de Crotálidos/química , Venenos de Crotálidos/farmacología , Metaloendopeptidasas/química , Metaloendopeptidasas/farmacología , Neovascularización Fisiológica/efectos de los fármacos , Venenos de Serpiente/enzimología , Animales , Biomarcadores/metabolismo , Fibrosis/inducido químicamente , Fibrosis/metabolismo , Hemoglobinas/metabolismo , Inflamación/inducido químicamente , Masculino , Ratones , Dominios Proteicos , Factor A de Crecimiento Endotelial Vascular/metabolismo , Veneno de Bothrops JararacaRESUMEN
The present work reports the effects of a C-type lectin (BpLec) isolated from Bothrops pauloensis snake venom upon in vitro and in vivo angiogenesis models. Initially, we noted that BpLec was not cytotoxic to endothelial cells (tEnd) in doses up to 40µg/mL, but lower doses (2.5µg/mL, 5µg/mL, 10µg/mL and 20µg/mL) reduced tEnd cells adhesion to some extracellular matrix proteins and inhibited the in vitro vessel formation in Matrigel assay stimulated by bFGF. ß-galactosides (d-lactose, N-acetyl-d-galactosamine and d-galactose) at 400mM reversed the effect of BpLec on tEnd cells adhesion, whereas d-galactose (400mM) partially reversed BpLec property of inhibiting vessel formation by tEnd cells in Matrigel. In vivo assays showed that BpLec increased hemoglobin content and capillary vessels number in polyether-polyurethane sponge discs subcutaneously implanted into dorsal skin mice. Additionally, BpLec also reduced collagen deposition and did not induce a pro-inflammatory response, as demonstrated by the decreased the secretion of some inflammatory cytokines, whereas myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) activities were not altered by BpLec. Taken together, our results indicate that BpLec might represent an interesting angiogenesis and inflammatory modulator that could also be used for searching possible therapeutic targets involved in these processes.
Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Bothrops , Venenos de Crotálidos/química , Lectinas Tipo C/metabolismo , Acetilglucosaminidasa/metabolismo , Inhibidores de la Angiogénesis/aislamiento & purificación , Inhibidores de la Angiogénesis/toxicidad , Animales , Adhesión Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Lectinas Tipo C/aislamiento & purificación , Masculino , Ratones , Ratones Endogámicos BALB C , Neovascularización Fisiológica/efectos de los fármacos , Peroxidasa/metabolismoRESUMEN
BACKGROUND: TNF-α is a key cytokine involved in prostate carcinogenesis and is mediated by the TNF-α receptor type 1 (TNFR-1). This receptor triggers two opposite pathways: cell death or cell survival and presents a protective or stimulator role in cancer. Thus, the purpose of this study was to evaluate the role of TNF signaling in chemically induced prostate carcinogenesis in mice. METHODS: C57bl/6 wild type (WT) and p55 TNFR-1 knockout mice (KO) were treated with mineral oil (control) or N-methyl N-nitrosurea (MNU) in association with testosterone (MNU+T, single injection of 40 mg/kg and weekly injection 2 mg/kg, respectively) over the course of 6 months. After this induction period, prostate samples were processed for histological and biochemical analysis. RESULTS: MNU+T treatment led to the development of prostate intraepithelial neoplasia (PIN) and adenocarcinoma (PCa) in both WT and KO animals; however, the incidence of PCa was lower in KO group than in WT. Cell proliferation analysis showed that PCNA levels were significantly lower in the KO group, even after carcinogenesis induction. Furthermore, the prostate of KO animals had lower levels of p65 and p-mTOR after treatment with MNU+T than WT. There was also a decrease in prostate androgen receptor levels after induction of carcinogenesis in both KO and WT mice. Regarding the extracellular matrix in the prostate, KO mice had higher levels of fibronectin and lower levels of matrix metalloproteinase 2 (MMP2) after carcinogenesis. Finally, there was a similar increase in apoptosis in both groups after carcinogenesis, indicating that the TNAFr1 pathway in prostate carcinogenesis presented proliferative, and not apoptotic, stimuli. CONCLUSIONS: TNF-α, through its receptor TNFR-1, promoted cell proliferation and cell survival in prostate by activation of the AKT/mTOR and NFKB pathway, which stimulated prostate carcinogenesis in chemically induced mice. Prostate 76: 917-926, 2016. © 2016 Wiley Periodicals, Inc.
Asunto(s)
Carcinogénesis , Neoplasias de la Próstata , Receptores Tipo I de Factores de Necrosis Tumoral/fisiología , Factor de Necrosis Tumoral alfa/fisiología , Adenocarcinoma/patología , Animales , Apoptosis , Carcinogénesis/patología , Proliferación Celular , Supervivencia Celular , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , FN-kappa B/metabolismo , Antígeno Nuclear de Célula en Proliferación/análisis , Neoplasia Intraepitelial Prostática/patología , Neoplasias de la Próstata/química , Neoplasias de la Próstata/patología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptores Androgénicos/análisis , Receptores Tipo I de Factores de Necrosis Tumoral/deficiencia , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Serina-Treonina Quinasas TOR/análisis , Serina-Treonina Quinasas TOR/metabolismo , Factor de Transcripción ReIA/análisisRESUMEN
Chagas disease, which is caused by the parasite Trypanosoma cruzi, is an important cause of cardiomyopathy in Latin America. It is estimated that 10%-30% of all infected individuals will acquire chronic chagasic cardiomyopathy (CCC). The etiology of CCC is multifactorial and involves parasite genotype, host genetic polymorphisms, immune response, signaling pathways and autoimmune progression. Herein we verified the impact of the recombinant form of P21 (rP21), a secreted T. cruzi protein involved in host cell invasion, on progression of inflammatory process in a polyester sponge-induced inflammation model. Results indicated that rP21 can recruit immune cells induce myeloperoxidase and IL-4 production and decrease blood vessels formation compared to controls in vitro and in vivo. In conclusion, T. cruzi P21 may be a potential target for the development of P21 antagonist compounds to treat chagasic cardiomyopathy.
Asunto(s)
Cardiomiopatías/etiología , Enfermedad de Chagas/patología , Proteínas Protozoarias/antagonistas & inhibidores , Trypanosoma cruzi/metabolismo , Animales , Cardiomiopatías/tratamiento farmacológico , Adhesión Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Enfermedad de Chagas/tratamiento farmacológico , Enfermedad de Chagas/parasitología , Quimiotaxis/efectos de los fármacos , Citocinas/metabolismo , Modelos Animales de Enfermedad , Inflamación/inducido químicamente , Inflamación/metabolismo , Inflamación/prevención & control , Interleucina-4/metabolismo , Leucocitos/citología , Leucocitos/inmunología , Leucocitos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Neovascularización Fisiológica/efectos de los fármacos , Peroxidasa/metabolismo , Proteínas Protozoarias/genética , Proteínas Protozoarias/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/farmacología , Proteínas Recombinantes/uso terapéutico , Trypanosoma cruzi/aislamiento & purificaciónRESUMEN
Integrins are involved in a number of physio-pathological processes including wound healing, chronic inflammation and neoplasias. Blocking its activity is potentially of therapeutic value in these conditions. We investigated whether DisBa-01, a recombinant His-tag RGD-disintegrin from Bothrops alternatus snake venom, could modulate key events (inflammatory cell recruitment/activation, neovascularization and extracellular matrix deposition) of the proliferative fibrovascular tissue induced by polyether polyurethane sponge implants in mice. The hemoglobin content (µg/mg wet tissue), blood flow measurements (laser Doppler perfusion imaging) and number of vessels in the implants, used as indices of vascularization, showed that the disintegrin dose-dependently reduced angiogenesis in the implants relative to the Saline-treated group. DisBa-01 inhibited neutrophil and macrophage content as determined by the myeloperoxidase (MPO) and N-acetyl-ß-D-glucosaminidase (NAG) activities, respectively. Similarly, down regulation of the fibrogenic component studied (collagen deposition) was observed in DisBa-01-treated implants. VEGF, bFGF, TNF-α, CXCL1 and CCL2 levels were also decreased by the disintegrin. The inhibitory effect of this αvß3-blocking disintegrin on the angiogenic, inflammatory, and fibrogenic components of the fibrovascular tissue induced by the synthetic matrix extends the range of DisBa-01 actions and may indicate its therapeutic potential in controlling angiogenesis in fibroproliferative diseases.
Asunto(s)
Bothrops/metabolismo , Venenos de Crotálidos/análisis , Desintegrinas/farmacología , Matriz Extracelular/efectos de los fármacos , Inflamación/prevención & control , Neovascularización Fisiológica/efectos de los fármacos , Proteínas Recombinantes/farmacología , Acetilglucosaminidasa/metabolismo , Animales , Venenos de Crotálidos/farmacología , Desintegrinas/análisis , Evaluación Preclínica de Medicamentos , Flujometría por Láser-Doppler , Macrófagos/efectos de los fármacos , Ratones , Peroxidasa/metabolismo , Poliuretanos , Flujo Sanguíneo Regional/efectos de los fármacosRESUMEN
A relationship between compromised muscles and other tissues has been demonstrated in mdx mouse, an animal model studied for understanding of Duchenne muscular dystrophy. The hypothesis is that changes in the calcaneal tendon of mdx mice occur previous to the onset of rigorous and most marked episodes of muscle degeneration, which start suddenly after 21 days of life. Thus, this study aimed to identify possible alterations in the calcaneal tendon of mdx mouse at 21 days of age. Control and mdx tendons were submitted to mechanical tensile testing, quantification of hydroxyproline, and staining with toluidine blue and picrosirius red. Hydroxyproline content was similar between mdx and control groups. The control tendon presented higher mechanical strength (load, stress, and elastic modulus) and its morphological analysis showed a larger number of round fibroblasts, nuclei with well-decondensed chromatin, and slightly metachromatic well-stained cytoplasmic material, different from that observed in mdx tendons. The results suggest that the absence of dystrophin in mdx mouse can provoke directly or indirectly alterations in the mechanical properties and morphology of the calcaneal tendon.
Asunto(s)
Calcáneo/anatomía & histología , Modelos Animales de Enfermedad , Músculo Esquelético/anatomía & histología , Distrofias Musculares/patología , Estrés Mecánico , Tendones/anatomía & histología , Animales , Fenómenos Biomecánicos , Calcáneo/metabolismo , Hidroxiprolina/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos mdx , Músculo Esquelético/metabolismo , Distrofias Musculares/metabolismo , Tendones/metabolismo , Resistencia a la TracciónRESUMEN
Several studies have demonstrated the relationship between exercise and the extracellular matrix of muscle tendons, and have described alterations in their structural and biochemical properties when subjected to strenuous exercise. However, little is known about what happens to tendons when they are subjected to stretching. We evaluated the changes in the composition and structure of rat calcaneal tendons subjected to a stretching program. The animals had their muscles stretched for 30 s with 30 s of rest, with 10 repetitions, three and five times a week for 21 days. For morphological analysis, the sections were stained with hematoxylin-eosin and toluidine blue. For biochemical analysis, the tendons were treated with 4 M guanidine hydrochloride and analyzed in SDS-PAGE. The contents of total proteins and glycosaminoglycans were also measured. In the sections stained with toluidine blue, we could observe an increase of rounded cells, especially in the enthesis region. In the region next to the enthesis was a metachromatic region, which was more intensely stained in the stretched groups. In the tension regions, the cells appeared more aligned. Cellularity increased in both regions. The SDS-PAGE analysis showed a larger amount of collagen in the stretched groups and a polydispersed component of 65 kDa in all the groups. The amounts of proteins and glycosaminoglycans were also larger in the stretched tendons. The agarose-gel electrophoresis confirmed the presence of dermatan sulfate in the tension and compression regions, and of chondroitin sulfate only in the latter. Our results showed that the stretching stimulus changed the cellularity and the amount of the extracellular matrix compounds, confirming that tendons are dynamic structures with a capacity to detect alterations in their load.
Asunto(s)
Sulfatos de Condroitina/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Matriz Extracelular/metabolismo , Ejercicios de Estiramiento Muscular , Condicionamiento Físico Animal , Tendones/metabolismo , Animales , Ratas , Ratas WistarRESUMEN
The effect of mesterolone and intensive treadmill training (6 weeks, 5 days/week, means: 15.82 m/min and 45.8 min/day) in Achilles tendon remodeling was evaluated. Sedentary mice treated with mesterolone (Sed-M) or vehicle (Sed-C, placebo/control) and corresponding exercised (Ex-M and Ex-C) were examined. SDS-polyacrylamide gel electrophoresis was used for determining collagen bands and hydroxyproline concentration. Collagen fibril diameter, the area and number of fibrils contained in an area probe, and the ultrastructure of fibroblasts (tenocytes) were determined. The presence of collagen was notable in the tendons of all groups. Collagen alpha(1/)alpha(2) bands in Sed-M, Ex-C, and Ex-M were higher than in Sed-C, as shown by hydroxyproline content, but collagen beta-chain appeared only in Ex-C. Noticeable bands of non-collagenous proteins were found in Sed-M and Ex-M. The number of fibrils in the area probe increased markedly in Sed-M and Ex-C (12-fold), but their diameter and area were unchanged compared with Sed-C. In Ex-M, the fibril number decreased by three-fold to 3.5-fold compared with Sed-M and Ex-C, whereas diameter and area increased. Sed-C tenocytes appeared quiescent, whereas those in the other groups seemed to be engaged in protein synthesis. The density of tenocytes was smaller in Sed-C than in Ex-C, Sed-M, and Ex-M. Thus, mechanical stimuli and mesterolone alter the morphology of tenocytes and the composition of the tendon, probably through fibrillogenesis and/or increased intermolecular cross-links. The ergogenic effect is evidenced by the activation of collagenous and non-collagenous protein synthesis and the increase in the diameter and area of collagen fibrils. This study might be relevant to clinical sports medicine.
Asunto(s)
Tendón Calcáneo/fisiología , Mesterolona/farmacología , Condicionamiento Físico Animal , Regeneración , Tendón Calcáneo/citología , Tendón Calcáneo/efectos de los fármacos , Animales , Colágenos Fibrilares/metabolismo , Fibroblastos/ultraestructura , Hidroxiprolina/metabolismo , Ratones , Ratones TransgénicosRESUMEN
The role of physical activity in affecting the composition of extracellular matrix and mechanical properties of tendons has been well studied, but little is known about the role of passive stretching. The purpose of this study was to test the hypothesis that stimulation by passive stretching may change the composition and mechanical properties of tendons. Three-month-old Wistar rats were divided into three groups: the control, animals were not submitted to stretching procedures; groups that had their calcaneal tendons manually stretched three or five times a week, for 21 days. Afterward, the calcaneal tendons were removed and assayed for hydroxyproline content and biomechanical test. The hydroxyproline content in the stretched groups was higher, suggesting that more collagen was present in the tendons of these groups. These tendons also showed higher values of maximum stress and modulus of elasticity or Young's modulus. These results indicate that stretching leads to alterations in the synthesis of the extracellular matrix components and in the mechanical properties of tendons.