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1.
Hernia ; 24(6): 1325-1336, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32306141

RESUMO

PURPOSE: To evaluate the vas deferens and testicles of rats submitted to bilateral inguinotomy and polypropylene (PP) mesh placement. METHOD: Sixty Wistar rats were randomized into three groups: Control (inguinotomy only), mesh placement over the vas deferens (Mesh-DD) or under the spermatic funiculus (Mesh-SF). The following analyses were performed: vas deferens morphometry (lumen area and wall thickness), quantification of collagen fibers, spermatogenesis, apoptosis (cleaved caspase-3 and TUNEL) and cellular proliferation (Ki67). Quantitative gene expression (qPCR) for apoptosis and inflammatory cytokines were evaluated by RT-PCR. RESULTS: In the apoptosis pathway, Mesh-DD showed one upregulated gene (Il10) and three downregulated genes (Fadd, Tnfrsf1b and Xiap). In Mesh-SF, 17 genes were downregulated. In the inflammation pathway (Mesh-DD), one gene was upregulated (Il1r1), and one gene was downregulated (Ccl12). In Mesh-SF, three genes were upregulated (Il1r1, Tnfsf13b and Csf1), and two were downregulated (Ccl12 and Csf2). PP mesh placement preserved spermatogenesis and did not alter the vas deferens or the testicle. In the ductus deferens, there was reduced luminal area (30 days), increased wall thickness (90 days), and increased type III collagen and cell proliferation (30 and 90 days) (p < 0.05). In the testicle, cell proliferation was greater in the Mesh-DD (p < 0.05). CONCLUSIONS: PP mesh, whether or not in direct contact with spermatic funicular structures, induces changes that were not sufficient to cause damage to the evaluated organs.


Assuntos
Genitália/cirurgia , Polipropilenos/uso terapêutico , Telas Cirúrgicas/normas , Animais , Modelos Animais de Doenças , Masculino , Ratos , Ratos Wistar
3.
J Proteomics ; 151: 162-173, 2017 01 16.
Artigo em Inglês | MEDLINE | ID: mdl-27208787

RESUMO

Intestinal ischemia and reperfusion injury are widely used models, which result into tissue injury and multiple organ failure also observed after trauma and surgery. Ischemic preconditioning (IPC) preceding ischemia and reperfusion (IR) was shown to attenuate this injury and has a potential therapeutic application; however the exact underlying mechanism is not clear. Neutrophils play an important role in the mechanism of injuries caused by ischemia and reperfusion while IPC led to a decrease in neutrophil stimulation and activation. The effect of preconditioning on the neutrophil proteome is unclear. Proteomic analysis has been ratified as an appropriate tool for studying complex systems. In order to evaluate the effect of IPC preceding 45min of ischemia on the proteome of neutrophils we used Wistar rats divided in four experimental groups: Control, sham laparotomy, intestinal ischemia reperfusion and ischemic preconditioning. After neutrophil separation, proteins were extracted, trypsin digested and the resulting peptides were iTRAQ labeled followed by HILIC fractionation and nLC-MS/MS analysis. After database searches, normalization and statistical analysis our proteomic analysis resulted in the identification of 2437 protein groups that were assigned to five different clusters based on the relative abundance profiles among the experimental groups. The clustering followed by statistical analysis led to the identification of significantly up and downregulated proteins in IR and IPC. Cluster based KEGG pathways analysis revealed up- regulation of actin cytoskeleton, metabolism, Fc gamma R mediated phagocytosis, chemokine signaling, focal adhesion and leukocyte transendothelial migration whereas downregulation in ribosome, spliceosome, RNA transport, protein processing in endoplasmic reticulum and proteasome, after intestinal ischemic preconditioning. Furthermore, enzyme prediction analysis revealed the regulation of some important antioxidant enzymes and having their role in reactive oxygen species production. To our knowledge, this work describes the most comprehensive and detailed quantitative proteomic study of the neutrophil showing the beneficial role of ischemic preconditioning and its effects on the neutrophil proteome. This data will be helpful to understand the effect of underlying protective mechanisms modulating the role of PMNs after IPC and provide a trustworthy basis for future studies. BIOLOGICAL SIGNIFICANCE: Preconditioning is a relevant strategy to overcome clinical implications from ischemia and reperfusion. Such implications have the neutrophil as a major player. Although many publications describe specific biochemical and physiological roles of the neutrophil in such conditions, there is no report of a proteomic study providing a broader view of this scenario. Here we describe a group of proteins significantly regulated by ischemia and reperfusion being such regulation prevented by preconditioning. Such finding may provide relevant information for a deeper understanding of the mechanisms involved, as well as serve as basis for future biomarker or drug target assays.


Assuntos
Precondicionamento Isquêmico , Neutrófilos/química , Proteoma/análise , Proteômica/métodos , Animais , Movimento Celular , Análise por Conglomerados , Intestinos/patologia , Oxirredutases , Ratos , Ratos Wistar , Traumatismo por Reperfusão , Proteínas Ribossômicas/análise
4.
Transplant Proc ; 46(6): 1862-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25131055

RESUMO

The aim of this study was evaluate the beta blocker atenolol (AT) and ischemic preconditioning (IPC) strategies for tissue protection against systemic effects of intestinal ischemia (I) and reperfusion (R) injury. Forty-two rats were pretreated with AT (1.5 mg · kg(-1)), 0.9% saline solution (SS; 0.1 mL), or IPC and then subjected to prolonged occlusion of the superior mesenteric artery for 60 minutes leading to I followed or not by 120 minutes of R, according to the group. For IPC, 5 minutes of I prior to 10 minutes of R were established. After this process of I or I-R, the right lung of each animal was adequately prepared for staining with hematoxylin and eosin and subsequent histologic analysis for quantification of inflammatory infiltrate was done. The left lung was frozen and prepared for assessment of oxidative stress by the quantification of thiobarbituric acid-reactivity substances (TBARS). Histologic analysis showed an important inflammatory infiltrate in the I-R + SS (I-R + SS = 4.5), which was significantly (P < .05) reduced by IPC (I-R + IPC = 3.0) or AT (I-R + AT = 3.0). Likewise, the TBARS levels were decreased by both strategies (I-R + SS = 0.63; I-R + IPC = 0.23; I-R + AT = 0.38; P < .05). Our results showed that AT and IPC attenuate pulmonary lesions caused by intestinal I and R process.


Assuntos
Atenolol/farmacologia , Intestinos/irrigação sanguínea , Precondicionamento Isquêmico/métodos , Lesão Pulmonar/prevenção & controle , Pulmão/irrigação sanguínea , Traumatismo por Reperfusão/prevenção & controle , Antagonistas de Receptores Adrenérgicos beta 1/farmacologia , Animais , Modelos Animais de Doenças , Lesão Pulmonar/etiologia , Masculino , Ratos , Ratos Wistar , Traumatismo por Reperfusão/complicações
5.
Transplant Proc ; 46(6): 1852-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25131053

RESUMO

To study the role of heparin and ischemic preconditioning (IPC) in cardiac injury after intestinal ischemia (I) and reperfusion (R), 54 rats underwent 60 minutes of I, which was produced by occlusion of the superior mesenteric artery, and/or 120 minutes of R. The IPC group had the I procedure stimulation for 5 minutes and R for 10 minutes. The control group was subjected to sham surgery only, and the other groups were injected with saline solution (SS; 0.1 mL) or heparin (100 IU/kg) via the inferior cava vein 5 minutes before I and 5 minutes before R and 55 minutes after the R begins in I-R groups. In all animals, cardiac samples were stained with hematoxylin and eosin for optical microscopy analysis, and other sample was processed for lipid peroxidation determination. In I-R groups, both heparin and IPC showed significant protection compared to the SS group; conversely, in animals subjected only to I, no protection was observed. Moreover, when heparin was associated with IPC, I-R protection was compromised and the ischemic injury increased. Data showed that IPC and heparin attenuated cardiac dysfunction caused by intestinal I and I-R, but when used in association did not show beneficial effects.


Assuntos
Cardiomiopatias/prevenção & controle , Heparina/uso terapêutico , Intestinos/irrigação sanguínea , Precondicionamento Isquêmico/métodos , Traumatismo por Reperfusão/prevenção & controle , Animais , Anticoagulantes/uso terapêutico , Cardiomiopatias/etiologia , Modelos Animais de Doenças , Masculino , Ratos , Ratos Wistar , Traumatismo por Reperfusão/complicações
6.
Chir Main ; 32(4): 206-9, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23867724

RESUMO

Surgery to transfer the axillary nerve and the nerve of the long head of the triceps presents two obstacles: 1) the access portals are not standardized and 2) the nerves are for their larger part approached through large incisions. The goal of this study was to explore the feasibility of an endoscopic microsurgical approach. The posterior aspect of a cadaver shoulder was approached through three communicating mini-incisions. The Da Vinci robot camera was installed on a central trocart, and the instrument arms on the adjacent trocarts. A gas insufflation distended the soft tissues up to the lateral axillary space. The branches of the axillary nerve and the nerve to the long head of the triceps brachii muscle were identified. The dissection of the axillary nerve trunk and its branches was easy. The posterior humeral circumflex veins and artery were dissected as well without any difficulty. Finding the nerve to the long head of the triceps brachii was found to be more challenging because of its deeper location. Robots properties allow performing conventional microsurgery: elimination of the physiologic tremor and multiplication of the movements. They also facilitate the endoscopic approach of the peripheral nerves, as seen in our results on the terminal branches of the axillary nerve and the nerve to the long head of the triceps brachii.


Assuntos
Axila/inervação , Plexo Braquial/cirurgia , Microcirurgia/métodos , Robótica , Axila/cirurgia , Cadáver , Estudos de Viabilidade , Humanos
7.
Transplant Proc ; 44(8): 2321-5, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23026584

RESUMO

BACKGROUND: Dysfunction of the liver after transplantation may be related to the graft size and ischemia/reperfusion (I/R) injury. N-Acetylcysteine (NAC) exerts beneficial effects on livers undergoing ischemia reperfusion. We sought to evaluate NAC modulation on reduced livers associated with I/R injury. METHODS: Male C57BL/6 mice of 8 weeks of age were divided into groups: 50% hepatectomy (G-Hep); NAC (G-Hep + NAC [150 mg/kg]) via vena cava 15 minutes before hepatectomy; ischemia (G-Hep + IR); NAC with hepatectomy (G-IR + Hep + Nac); and IR using 30 minutes selective hepatic occlusion and reperfusion for 24 hours. After 24 hours, the remaining liver was removed, for staining with hematoxylin and eosin or labeling by proliferating cell nuclear antigen. Blood was collected for biochemical evaluations. Significance was considered for P ≤ .05. RESULTS: Aspartate aminotransferase was high in all studied groups reflecting the hepatectomy and intervention. injuries. However, when assessing alanine aminotransferase, which depicts liver function, induction of IR promoted a greater increase than hepatectomy (P = .0003). NAC decreased ALT activity in all groups, even in association with I/R (P < .05), reflecting a modulation of the injury. Necrosis resulting from IR was mitigated by NAC. The experimental model of 50% reduced live promoted regeneration of the hepatic remnant, which was accentuated by NAC, according to the total number of hepatocytes and PCNA values. CONCLUSION: NAC preserved the remnant liver in mice and stimulates regeneration even after IR injury.


Assuntos
Acetilcisteína/farmacologia , Proliferação de Células/efeitos dos fármacos , Hepatectomia , Regeneração Hepática/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/cirurgia , Traumatismo por Reperfusão/tratamento farmacológico , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Biomarcadores/sangue , Biomarcadores/metabolismo , Citoproteção , Modelos Animais de Doenças , Imuno-Histoquímica , Fígado/irrigação sanguínea , Fígado/metabolismo , Fígado/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Necrose , Antígeno Nuclear de Célula em Proliferação/metabolismo , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologia , Fatores de Tempo
8.
Transplant Proc ; 41(3): 827-9, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19376363

RESUMO

PURPOSE: The purpose of this study was to establish morphologically the best time of vascular occlusion to induce ischemic preconditioning (IPC) for rat small bowel undergoing ischemia and reperfusion injury. METHODS: After approval by the Ethics Committee, 36 EPM-1 young adult Wistar rats from 300-350 g were distributed into 6 groups: sham (S); ischemia and reperfusion (IR), with 50 minutes of cranial mesenteric artery occlusion and 30 minutes of reperfusion; IPC with 1 cycle of 2 minutes (IPC-2), 5 minutes (IPC-5), 10 minutes (IPC-10), or 15 minutes (IPC-15), followed by sustained IR. The animals anesthetized with ketamine (60 mg/kg) and xylazine (10 mg/kg) intramuscular (IM), were maintained on mattress heat, hydrated with saline (80 mL/kg), and injected with 100 IU heparin. Samples of jejunum were stained with hematoxylin and eosin (HE) and classified according to Park et al. Statistical analysis of results was performed using Kruskal-Wallis tests (P < .05). RESULTS: The histological evaluation showed no difference between IR and IPC15 rats (5.2 and 5, respectively; P = .84). Greater jejunal injury was observed with IPC15 (5) compared with other groups (IPC2 = 3, P = .03; IPC5 = 3.2, P = .05; IPC10 = 2.8, P = .02, respectively). There was no difference between groups IPC2 x IPC5 x IPC10. CONCLUSION: Morphologically, IPC with short times promoted greater intestinal protection against the IR lesion in rats.


Assuntos
Intestino Delgado/fisiopatologia , Precondicionamento Isquêmico/métodos , Traumatismo por Reperfusão/fisiopatologia , Animais , Mucosa Intestinal/lesões , Mucosa Intestinal/patologia , Intestino Delgado/patologia , Isquemia , Masculino , Ratos , Ratos Wistar , Reperfusão , Traumatismo por Reperfusão/patologia , Traumatismo por Reperfusão/prevenção & controle
9.
Transplant Proc ; 41(3): 830-3, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19376364

RESUMO

PURPOSE: The purpose of this study was to evaluate estradiol serum levels and follicular development in rats subjected to ovarian autologous transplantation with or without remote ischemic preconditioning (R-IPC). METHODS: Seventy-two adult female Wistar EPM-1 rats were distributed into 3 groups: (1) controls; (2) ovarian transplantation; and (3) ovarian transplantation + R-IPC. The groups were divided into subgroups, according to the prefixed date for euthanasia: 24 hours, 48 hours, 72 hours, and 7th postoperative day (PO). R-IPC was performed by clamping the common iliac artery for a 15-minute period of ischemia followed by 15 minutes of reperfusion, before undergoing ovarian transplantation. The graft was fixed to the retroperitoneum with a simple 8-0 prolene thread. Blood samples were collected from the vena cava. For evaluation of follicular development, the ovarian follicles were classified as immature and mature follicles besides corpora lutea. Only the viable follicles and functioning corpora lutea were counted. RESULTS: At 72 hours, the R-IPC group showed higher estradiol values than the other groups, which were similar. After 24 hours the mean values were similar among all groups, and at 48 hours the R-IPC group was similar to the transplanted group without IPC. Animals undergoing R-IPC showed superior morphologic aspects, but 7 days after transplantation the morphology was worse in all groups. R-IPC enhanced the number of immature follicles at 48 hours (P > .05) and number of mature follicles from 24 hours to 48 hours after transplantation (P < .01). Functioning corpora lutea number was increased as well. CONCLUSION: R-IPC increased the estradiol levels in autologous ovarian transplants associated with better graft morphology and more mature follicles.


Assuntos
Estradiol/sangue , Precondicionamento Isquêmico/métodos , Folículo Ovariano/fisiologia , Ovário/transplante , Animais , Corpo Lúteo/patologia , Corpo Lúteo/fisiologia , Feminino , Inflamação/patologia , Necrose , Folículo Ovariano/patologia , Ovariectomia , Ovário/patologia , Ratos , Ratos Wistar , Valores de Referência , Transplante Isogênico/patologia
10.
Transplant Proc ; 40(3): 861-4, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18455037

RESUMO

PURPOSE: Verify the optimum remote vascular occlusion time to reduce ovarian injury in autologous transplants in rats. METHODS: Twenty-four adult female rats were assigned to four groups: GC (control group): bilateral oophorectomy followed by ovary transplant; GIPC (ischemic preconditioning group): remote ischemic preconditioning at the iliac artery for 5, 10, and 15 minutes (GIPC-5, GIPC-20, and GIPC-15) previous to bilateral oophorectomy and ovarian transplantation. The right ovary was fixed in the retroperitoneum. Euthanasia was performed 4 days after the surgical procedure. The follicles were counted and classified as developing versus atretic. The immunohistochemical assay identified vascular factor of endothelial growth (VEGF) in the ovarian stroma and assessed the proliferation capacity by means of the Ki-67 in the ovarian follicles. RESULTS: Every group showed an inflammatory infiltrate, luteous body, and ovarian follicles in several phases of development. The ischemic preconditioning groups displayed greater amounts of viable ovarian follicles and increased vascularization and vasodilatation than the control group. GIPC-15 showed the highest amount of viable follicles compared to the others (P < .05 GIPC-15 vs GC; GIPC-15 vs GIPC-5). More VEGF-labeled cells were observed in GIPC-10 than the control group (P < .05, GIPC-10 vs GC). The proliferation index assessed by Ki-67 marking showed GC: 80%; GIPC-5: 76%; GIPC-10: 67%; and GIPC-15: 64% (P > .05). CONCLUSIONS: The PCI-15 cohort seem to be the most adequate timing to achieve functional support and preservation of a greater number of viable ovarian follicles.


Assuntos
Precondicionamento Isquêmico/métodos , Folículo Ovariano/citologia , Ovário/transplante , Animais , Feminino , Modelos Animais , Folículo Ovariano/irrigação sanguínea , Ovariectomia , Ratos , Ratos Wistar , Transplante Autólogo
11.
Transplant Proc ; 37(1): 425-7, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15808665

RESUMO

AIM: To study the effects of N-acetylcysteine and ischemic preconditioning on the portal triad clamping compared to arterial and portal clamping alone. METHODS: Eighty EPM 1-Wistar rats were randomized into two groups, depending on inclusion (Group 1) or not (Group 2) of the bile duct in the hepatic vascular pedicle occlusion. Each group was divided into four subgroups as follows. IR 1: 20 minutes after celiotomy, the pedicle containing vascular elements and bile duct to the left lateral and median liver lobes was occluded for 40 minutes, followed by 30 minutes of reperfusion. IPC 1: after 10 minutes of ischemia and 10 minutes of reperfusion, the ischemic preconditioning period, the rats were submitted to the same procedure described for IR 1 Group. NAC 1: the rats received N-acetylcysteine (150 mg/kg) 15 minutes before 40 minutes of ischemia and 5 minutes before 30 minutes of reperfusion. SHAM 1: The hepatic pedicle for the lateral and median liver lobes was dissected after 20 minutes, the bile duct alone was clamped for 40 minutes, and released for an additional 30 minutes. In the IR 2, IPC 2, and NAC 2 groups, ischemia was achieved with an exclusive vascular occlusion. SHAM 2: dissection and observation for 90 minutes. The blood was sampled for liver enzyme levels. Statistical analysis was done (P

Assuntos
Acetilcisteína/farmacologia , Ductos Biliares/fisiologia , Precondicionamento Isquêmico , Circulação Hepática/fisiologia , Traumatismo por Reperfusão/prevenção & controle , Alanina Transaminase/metabolismo , Fosfatase Alcalina/metabolismo , Animais , Aspartato Aminotransferases/metabolismo , Bilirrubina/metabolismo , Circulação Hepática/efeitos dos fármacos , Masculino , Sistema Porta , Ratos , Ratos Wistar , gama-Glutamiltransferase/metabolismo
12.
Transplant Proc ; 36(4): 846-8, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15194290

RESUMO

This article seeks to standardize an experimental model of liver ischemia-reperfusion in rats following hemorrhagic shock modulated by N-acetylcysteine (NAC). Twenty-seven adult Wistar rats were randomized into three groups: the HS-IR-Garm underwent hemorrhagic shock with selective hepatic ischemia followed by reperfusion; the HSIR + NAC-G, the same procedure plus NAC; and the control group, only venous catheterization. Blood was withdrawn for 10 minutes until MABP reached 35 mm Hg, which was maintained for 1 hour. The blood was then reinjected as required to maintain MABP at that level. Ringer's lactate solution was infused in a volume equivalent to three times the shed blood, over a period of 15 minutes. Half of the shed blood was reinfused over 5 minutes. HSIR + NAC-G received 150 mg/kg of NAC, during treatment of the shock, and again 10 minutes before reperfusion and continued for 30 minutes. Finally, both groups were subjected to 40 minutes of warm selective hepatic ischemia and reperfusion for 1 hour. Data were analyzed by nonparametric tests (P < or =.05). Liver enzyme levels were higher in HS-IR-G (DHL = 6094 +/- 1688, AST = 746 +/- 175, and ALT = 457 +/- 90) than in HSIR + NAC-G group (DHL = 2920 +/- 284, AST = 419 +/- 113, and ALT = 253 +/- 26). The values in the control group were lower than both experimental groups (DHL = 965 +/- 173, AST = 163 +/- 42, and ALT = 82 +/- 28). Our data showed that liver ischemia-reperfusion injury following hemorrhagic shock produces important hepatic damage and that NAC reduces injury in this rat model.


Assuntos
Acetilcisteína/uso terapêutico , Fígado/irrigação sanguínea , Traumatismo por Reperfusão/prevenção & controle , Choque Hemorrágico/prevenção & controle , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Modelos Animais de Doenças , L-Lactato Desidrogenase/sangue , Fígado/patologia , Testes de Função Hepática , Ratos , Ratos Wistar
13.
Transplant Proc ; 36(4): 860-2, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15194294

RESUMO

The objective of this study was to evaluate the effect of ischemic preconditioning upon lesions produced by ischemia-reperfusion of the small intestine. Thirty EPM-1 Wistar rats were randomly distributed into three groups: ischemic preconditioning (IPC; n = 12), ischemia-reperfusion (I/R; n = 12), and control (C; n = 6). Laparotomy permitted isolation of the mesenteric artery for clamping. The animals were heparinized and hydrated. IPC was induced by: 10 minutes of ischemia followed by 10 minutes of reperfusion and then 50 minutes ischemia followed by another 30 minutes reperfusion. Group I/R was submitted to the same protocol except for the 20 minutes of preconditioning. Group C animals underwent only laparotomy for 100 minutes. After reperfusion small intestine fragments were examined histologically. Blood samples were obtained to measure LDH and lactate prior to euthanasia. Lactate values were significantly lower in the IPC as compared to I/R group, 39 versus 67 mg/dL, respectively (P < or =.05). However, neither IPC (grade 3) lesions of the mucosa versus I/R (grade 4) nor LDH values (PCI = 680, I/R = 873 U/L) were statistically different. Thus No morphological evidence of protection was observed following ischemic preconditioning.


Assuntos
Intestino Delgado/patologia , Precondicionamento Isquêmico , Traumatismo por Reperfusão/prevenção & controle , Animais , Modelos Animais de Doenças , Intestino Delgado/irrigação sanguínea , Masculino , Ratos , Ratos Wistar , Valores de Referência , Traumatismo por Reperfusão/patologia
14.
Microsurgery ; 21(4): 170-2, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11494387

RESUMO

Previous studies demonstrated that some immunosuppressive agents inhibit arterial intimal hyperplasia. Our previous studies demonstrated that gangliosides (Gang) have an immunosuppressive effect on as well as an anti-inflammatory role in the wound-healing process. Therefore, we decided to examine the effect of Gang on intimal hyperplasia. Twenty Wistar isogenic rats received a transverse division of the anterior wall of the femoral artery, followed by suturing using mononylon 10-0 under surgical microscopy and were then divided into two groups: Gang group, 3 mg/kg per day of Gang, and control group, vehicle, intramuscularly from surgery to death (1 and 3 weeks, respectively). Concentric intimal hyperplasia was observed in arteries stained by hematoxylin-eosin in control and Gang groups. However, the media layer did not demonstrate any major alterations. After 3 weeks, the Gang group showed more intimal hyperplasia than the control group. Therefore, because intimal hyperplasia worsened in the presence of Gang after 3 weeks, further studies will be necessary to clarify its role in intimal proliferation.


Assuntos
Artéria Femoral/cirurgia , Displasia Fibromuscular/patologia , Gangliosídeos/farmacologia , Imunossupressores/farmacologia , Microcirurgia , Anastomose Cirúrgica , Animais , Artéria Femoral/patologia , Injeções Intramusculares , Masculino , Ratos , Ratos Wistar , Túnica Íntima/patologia , Túnica Média/patologia
16.
Acta cir. bras ; 15(supl.1): 7-9, 2000. tab, graf
Artigo em Inglês | LILACS | ID: lil-281309

RESUMO

Our previous study have demonstrated that Glycosphingolipids (GSLs) have an immunosuppressive effect on murine lymphoproliferation and IL-2 production. In the present study we examined the effect of a pool of Gangliosides (Gang) on spleen lymphocyte proliferation from either isogeneic strains of Wistar rats or BALB/c mice. Two hundred-fifty grams adult female isogeneic Wistar rats and 8-week-old BALB/c mice were used. The animals were sacrificed and the spleen harvested aseptically for cellular assays. Spleen cells suspensions were obtained by homogenization in RPMI 1640 with a loose tissue grinder. After washing, the cells were suspended in RPMI 1640 supplemented. Cell viability was measured by Trypan blue exclusion. Cells were cultured in triplicate using increasing concentrations of Gang (1; 2; 5; 10; 15; 20 mg/well) and in the presence of Concanavalin A. The cells were incubated for 48 hours and were pulsed with [3H] thymidine 18 hours prior to harvesting on glass fiber paper for counting in a beta-counter. Data were presented as rate of inhibition, as previously described. At concentrations 1 and 2 mug/well, Gang stimulated lymphoproliferation (30 percent and 50 percent, rats and mice respectively), while at concentration from 5 to 20 mg/well an increasing inhibition was observed for spleen cells from both mouse and rat (from 40 percent up to 80 percent). In preliminary studies we observed inhibition of mixed lymphocyte reaction on spleen cells from rats treated with Gang for 10 days (data not shown). Our data suggest that Gang may be investigated as a immunosuppressive drug in organ transplantation.


Assuntos
Animais , Feminino , Camundongos , Ratos , Baço/citologia , Gangliosídeos/farmacologia , Transtornos Linfoproliferativos , Adjuvantes Imunológicos , Glicoesfingolipídeos
17.
Acta cir. bras ; 15(supl.1): 10-2, 2000. ilus
Artigo em Português | LILACS | ID: lil-281310

RESUMO

Em trabalhos anteriores mostrou-se que os gangliosídeos (GSLs) têm um efeito inibitório sobre a proliferação linfocitária e a síntese de IL-2, assim como sobre a reação mista de linfócitos. Neste estudo objetivou-se avaliar o efeito dos GSLs sobre a resposta de hipersensibilidade retardada. Foram utilizados 12 camundongos BALB/c, machos, pesando em média 30 gramas, provenientes do biotério setorial da Disciplina de Parasitologia e mantidos por 5 dias para adaptação no biotério setorial da Disciplina de Técnica Operatória e Cirurgia Experimental da UNIFESP-EPM, recebendo água e ração própria para a espécie. Os animais foram distribuídos em três grupos, de acordo com as doses de GSLs, da seguinte forma: grupo 3mg. kg-1, grupo 9mg. kg-1 e grupo simulado (veículo). Os animais foram tratados, por via intramuscular, nos dias 0 e 4. O parâmetro avaliado foi o edema da pata traseira esquerda no local da inoculação do antígeno. Os animais foram anestesiados com Cetamina (60mg.kg-1) e Xilazina (10mg.kg-1), por via intramuscular, sendo em seguida submetidos à dissecção da veia jugular direita, por onde foram inoculadas 10(6) hemácias de Carneiro no dia 0, para sensibilização. No dia 4 subsequente, os animais foram novamente anestesiados e receberam, por via subcutânea, 10(8) hemácias de Carneiro, num volume de 0,02ml. Foram realizadas medidas do edema da pata traseira com paquímetro 24, 48, 72 e 96 horas após o desafio. Os dados mostraram que após 48h houve um aumento do edema em animais dos grupos simulado e 3mg (médias=2,3 and 2,1mm, respectivamente), e os camundongos do grupo 9mg não apresentaram aumento importante (média=0,1mm). Entretanto, após 72h, o grupo 9mg apresentou aumento de 1,7mm enquanto, os outros grupos não apresentaram mudança significativa no edema da pata (médias=0,2 e 0,8mm), grupos simulado e 3mg, respectivamente) comparados aos dados do dia antecedente. Após 96h, todos os grupos apresentaram desaparecimento do edema. Com base nos dados obtidos pode-se concluir que a resposta de hipersensibilidade retardada alterou-se na vigência de alta dose de GSLs.


Assuntos
Animais , Masculino , Camundongos , Gangliosídeos/efeitos adversos , Hipersensibilidade , Gangliosídeos/administração & dosagem , Camundongos Endogâmicos BALB C
18.
Acta cir. bras ; 15(supl.1): 13-5, 2000. graf
Artigo em Português | LILACS | ID: lil-281311

RESUMO

Imunomodulação mais específica e eficaz é uma meta importante a ser atingida na área de órgão. Neste sentido, foi estudado previamente o papel imunomodulador dos gangliosídeos "in vitro". No presente trabalho objetivou-se avaliar este efeito agora "in vivo", mimetizando a situação do transplante alogênico. Foram utilizados 26 ratos Wistar 1 EPM, machos, com 3 meses de idade, pesando cerca de 250g, procedentes do Centro de Desenvolvimento de Pesquisa Experimental em Medicina e Biologia. Os animais fora mantidos por 5 dias, para adaptação, no biotério setorial da Disciplina de Técnica Operatória e Cirurgia Experimental da UNIFESP-EPM, recebendo água e ração própria para a espécie. Os animais foram distribuídos em grupos conforme segue: grupos experimento (que receberam 1, 3 e 6 mg/kg/dia de gangliosídeos) e um grupo controle que recebeu veículo, todos por via intramuscular durante 7 dias consecutivos. No 8º dia, com os animais anestesiados com éter etílico foi feita a remoção cirúrgica do baço de todos os animais, os quais foram sacrificados por exsanguinação, ainda sob efeito anestésico. Os baços removidos foram processados para a obtenção de linfócitos os quais foram cultivados em placa de cultura com 96 poços, distribuídos da seguinte forma: 1,5x10(5) linfócitos viáveis de cada animal dos grupos experimento e controle foram cultivados com 1,5x10(5) linfócitos viáveis de um rato não tratado, sendo assim realizada a reação mista de linfócitos. Os linfócitos provenientes dos animais dos grupos controle e 1 mg apresentaram aumento da proliferação sem nenhuma alteração. Por outro lado, foi observada uma taxa de inibição ao redor de 70 por cento sobre a proliferação linfocitária dos animais dos grupos 3 e 6 mg comparados aos animais dos grupos controle e 1 mg. O resultado desta investigação estimula a utilização dos gangliosídeos no tratamento da rejeição alogênica.


Assuntos
Animais , Masculino , Ratos , Adjuvantes Imunológicos/uso terapêutico , Gangliosídeos/farmacologia , Baço/cirurgia , Contagem de Linfócitos/métodos , Glicoesfingolipídeos/farmacologia , Teste de Cultura Mista de Linfócitos/métodos
19.
Acta cir. bras ; 15(supl.1): 16-9, 2000. ilus, graf
Artigo em Português | LILACS | ID: lil-281312

RESUMO

Objetivo: Investigar o efeito dos gangliosídeos sobre a infiltração seqüencial de leucócitos e fibroblastos durante o processo de cicatrização usando um modelo de cicatrização da pele em ratos. Método: Foram utilizados 12 ratas EPM - 1 Wistar, com peso médio de 200 gramas e 4 meses de idade. Os animais procederam do Centro de Desenvolvimento de Pesquisa Experimental em Medicina e Biologia e foram mantidos por 5 dias para adaptação no biotério setorial da disciplina de Técnica Operatória e Cirurgia Experimental da UNIFESP-EPM, recebendo água e ração própria para a espécie. O protocolo anestésico utilizado foi uma associação de Cetamina (60mg.kg-1) e Xilazina (10mg.kg-1), por via intramuscular. Em seguida, realizava-se uma incisão longitudinal com 7 cm de extensão, na região dorsal paravertebral, interessando pele e tela subcutânea que foi fechada com pontos separados, com fio de prolene 7-0 e agulha triangular. As ratas foram distribuídas em dois grupos a saber: grupo experimento, que recebeu 3mg.kg-1.dia-1 de gangliosídeos, e um grupo controle, que recebeu veículo, ambos por via intramuscular durante 14 dias consecutivos. No 7º e 14º dias de pós-operatório foram ressecados fragmentos da pele e tela subcutânea para análise histológica, com a coloração de Tricrômio de Masson e Hematoxilina - Eosina. Resultados: As amostras apresentaram a mesma quantidade de colágeno em ambos os grupos mostrando que não houve inibição dos fibroblastos. Entretanto, a infiltração leucocitária foi retardada no grupo experimento quando comparado ao grupo controle. Conclusão: A alteração encontrada no processo cicatricial foi devida a um retardo na resposta inflamatória e não a uma inibição fibroblastos.


Assuntos
Animais , Feminino , Ratos , Cicatrização/fisiologia , Colágeno/fisiologia , Gangliosídeos/farmacologia , Inflamação , Ratos Wistar , Rejeição de Enxerto/prevenção & controle
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