Avaliamos a função cardíaca frente a Isquemia e reperfusão, em corações isolados de ratas intactas, com hipotireoidismo (Hipo), hipotireoidismo subclínico (HTS), redução dos níveis de hormônios ovarianos (OO) e a ambas condições (HTS+OO). Na estabilização, a Pressão Desenvolvida pelo Ventrículo Esquerdo (LVDP), Pressão Diastólica Final (EDP) e Pressão de Perfusão não diferiram. Porém, a Primeira Derivada Positiva das Pressões (+dP/dt), Primeira Derivada Negativa das Pressões (-dP/dt) e frequência cardíaca foram menores no grupo Hipo. Na reperfusão, os corações Hipo tiveram EDP, +dP/dt e -dP/dt semelhantes a estabilização. Assim, sugere-se que o HTS, a OO e o HTS + OO não alteraram a recuperação cardíaca após a isquemia. E que o Hipo é resistente à lesão por isquemia, apesar da função cardíaca deprimida. / We evaluate the cardiac function in relation to an ischemia and reperfusion event in hearts isolated from intact rats, With hypothyroidism (Hypo), subclinical hypothyroidism (HTS), decreased levels of ovarian hormones (OO) and both conditions (HTS + OO). At etabilization the Left Ventricle Developed Pressure (LVDP), Final Diastolic Pressure (EDP) and Perfusion Pressure did not differ. However, the First Positive Pressure Derivative (+ dP / dt), First Negative Pressure Derivative (-dP / dt) and heart rate were lower in the Hypo group. On reperfusion the hypothyroid hearts presented EDP, + dP / dt and -dP / dt Similar to stabilization. Thus, it is suggested that HTS, a OO e o HTS + OO Did not change cardiac recovery after ischemia.Is that hipo is resistant to ischemia injury, despite depressed cardiac function.

Aims There are few proven therapies that can protect against the inevitable ischaemia reperfusion injury (IRI) that occurs during renal transplantation. IRI increases the likelihood of delayed graft function (DGF), which negatively impacts on the long-term survival of a transplanted kidney. One enzyme of interest, heme oxygenase-1 (HO-1), degrades heme and protects against the oxidative stress that occurs secondary to IRI. Clinical renal recipients with higher HO-1 levels have improved graft function post transplant. Heme arginate (HA), a form of hemin, which has been used to treat porphyria for over 30 years, has repeatedly been shown to induce HO-1 in in vivo and in vitro macrophages. It is one of the few HO-1 inducers approved for clinical use and healthy volunteer studies confirmed that HA could also safely induce HO-1 in humans. Prior to the formal start of the MD, the University of Edinburgh successfully applied to NHS Blood and Transplant for funding to investigate whether giving HA to recipients of deceased donor renal grafts prior to transplant could upregulate HO-1 and whether this had any effect on the function and health of the grafts. This MD aims to explain the background behind the proposed study, the process of study approval, planning and trial logistics and protocol. This thesis then describes the methods of sample analysis, the results and future directions for the HOT (Heme Oxygenase-1 in renal Transplantation) study. Methods The HOT study planning and approval process took eight months and the first participant was randomised in January 2012. The study was sponsored by ACCORD, a joint company from University of Edinburgh and NHS Lothian, and recruited patients from the Edinburgh Royal Infirmary Transplant Unit. The protocol was followed to ensure that 40 recipients were randomised blind to either active (two doses 3mg kg-1 HA: pre-operatively, day 2) or placebo (NaCl: same schedule). To ensure that the primary outcome was fulfilled, recipient blood was taken daily for peripheral blood mononuclear cells (PBMC) extraction. After further blinding steps, the PBMCs were analysed for HO-1 protein and mRNA. The secondary outcome measures involved collecting urine for analysis of urinary biomarkers (KIM-1 and NGAL), taking renal graft biopsies pre-op and day 5 for renal HO-1 analysis and collecting renal function data. DGF was calculated daily. To ensure that all adverse event data was captured, the recipients were closely reviewed for 7 days and their renal function was monitored for 90 days. Results The final participant was recruited in May 2013 within the predicted timescale and to budget. This participant completed follow-up in August 2013. Of the 40 participants, three received the infusion but did not receive a transplant and therefore could not give primary outcome data. The remaining 37 did and this was analysed. Adverse events were equivalent between groups and there were no adverse reactions to HA. HA upregulated PBMC HO-1 protein at 24 hours compared to placebo: HA 11.1ng/ml (1.0- 37.0) vs. placebo 0.14ng/ml (-0.7- 0.3)(p= < 0.0001). PBMC HO-1 mRNA was also increased: HA 2.73 fold (1.8- 3.2) vs. placebo 1.41 fold (1.2- 2.2) (p=0.02). HA increased HO-1 protein immunopositivity in day 5 renal tissue compared with placebo: HA 0.21 (-24- 0.7) vs. placebo -0.03 (-76- 0.15) (p=0.02) and the percentage of HO-1 positive renal macrophages also increased: HA 50.8 cells per HPF (40.0- 59.8) vs. placebo 22.3 (0- 34.8) (p=0.012). Renal HO-1 mRNA was also increased in HA group: 2.02 (0.20- 4.03) fold increase compared to 1.68 (0.75- 10.39) fold in the placebo group but it was not significant (p= 0.451). Urinary biomarkers were reduced after HA but not significantly so. Histological injury and DGF rates were similar between the groups. Conclusion HA is safe and effective in renal transplant recipients as reported in this phase II, randomised, placebo controlled, blinded, single-centre study. The primary outcome was achieved and demonstrated for the first time that HA induces HO-1 in peripheral and renal macrophages in kidney transplant recipients. There was also evidence that HA increased HO-1 expression in renal tissue. There was no evidence that HA improved renal function or reduced injury as seen in animal models but it is recognised that the sample size was small and the study was not powered to these endpoints. Larger studies are planned to determine the impact of HO-1 upregulation on clinical outcomes and evaluate the benefit to patients at risk of IRI. The plans for HOT2 are expanded in this thesis.

A técnica de isquemia-reperfusão tem sido utilizada em cirurgias conservadoras do rim como a nefrectomia parcial e em transplantes renais. Para se realizar a isquemia pode-se fazer o bloqueio do fluxo sanguíneo da artéria renal ou o bloqueio simultâneo da artéria e da veia renal. O evento isquêmico acarreta em dano celular ao rim principalmente pelo estresse oxidativo local e a liberação de radicais livres assim como o aumento da resposta inflamatória. Diversos autores verificaram lesão renal após a isquemia-reperfusão, porém, apenas testes funcionais foram realizados até o momento. Os autores que tentaram avaliar a lesão morfológica do rim apenas fizeram a quantificação de escores subjetivos. O nosso objetivo é avaliar por quantificação estereológica o dano causado pela isquemia-reperfusão comparando o clampeamento somente arterial com o clampeamento arteriovenoso. Utilizamos 24 ratos wistar, machos, de quatro meses de idade. Os animais foram divididos em três grupos: o grupo Sham (n=8), o grupo de clampeamento somente da artéria renal (n=8) e o grupo de clampeamento simultâneo da artéria e da veia renal (n=8). Os animais foram submetidos a laparotomia mediana. Os animais do grupo Sham permaneceram os 60 minutos anestesiados mas sem obstrução do fluxo sanguíneo de seus vasos renais. Os animais do grupo de clampeamento arterial foram submetidos à clampeamento de sua arterial renal esquerda por 60 minutos e os animais do grupo de clampeamento arterial e venoso tiveram seus vasos renais esquerdos clampeados simultaneamente e em bloco pelo mesmo tempo. Após os 60 minutos os animais foram suturados e mantidos por 30 dias em caixas próprias sendo mortos por sobredose anestésica após decorrido esse tempo. Os rins foram coletados e mantidos em solução de formalina tamponada e posteriormente processados para análise histológica e estereológica. Foram analisados a densidade volumétrica (Vv) dos glomérulos, o número de glomérulos/mm3(Nv) e o volume glomerular médio (VGM). A Vv e Nv se encontrou reduzida nos rins esquerdos submetidos à isquemia mas foi somente significativa nos animais do grupo de clampeamento arterial e venoso. Mesmo usando o rato como modelo animal experimental, a partir de nossos resultados recomendamos o uso do clampeamento somente arterial nos casos em que mínina lesão ao rim é imperiosa. / The ischemia-reperfusion has been used in renal surgery such as conservative partial nephrectomy and kidney transplantation. To perform ischemia we can block the blood flow of renal artery or simultaneous blockade of the renal artery and vein. The ischemia leads to cellular damage to the kidney mainly by local oxidative stress and releasing free radicals as well as increased inflammatory response. Several authors related renal injury after ischemia-reperfusion, however; only functional tests were carried out to date. The authors attempted to evaluate the morphological lesion of the kidney using subjective scores. Our goal is assess by stereological quantification of the renal parenchyma after ischemia-reperfusion injury comparing arterial only clamping and arteriovenous clamping. We used 24 Wistar rats, males, four months old. The animals were divided into three groups: Sham group (n = 8), the group of clamping of the renal artery (n = 8) and the group of simultaneous clamping of renal artery and vein (n = 8). The animals underwent laparotomy. Sham group animals remained anesthetized for 60 minutes without blood flow obstruction of their renal vessels. The group animals underwent arterial clamping have had his left renal artery clamped for 60 minutes and the animals in the group of arterial and venous clamping had their left renal vessels clamped simultaneously for the same time. After 60 minutes the animals were sutured and kept for 30 days in their own boxes being killed by anesthetic overdose after that. The kidneys were collected and kept in formaldehyde solution and subsequently processed for histological and stereological analysis. We analyzed the volume density (Vv) of the glomeruli, the number of glomeruli/mm3 (Nv) and mean glomerular volume (MGV). The Vv and Nv was reduced in the ischemic left kidney but was only significant in animals of the group of arterial and venous clamping. Even using the rat as experimental animal model, from our results we recommend the use of arterial clamping only in cases where minimum injury to the kidney is imperative.

Made available in DSpace on 2014-06-11T19:31:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-22Bitstream added on 2014-06-13T19:40:58Z : No. of bitstreams: 1 francischetti_i_dr_botfm.pdf: 3036523 bytes, checksum: 16d5951b642a829416e73e1b1c9c6644 (MD5) / O uso da circulação extracorpórea desencadeia uma resposta inflamatória sistêmica, que somada às reações em cadeia relacionadas à isquemia-reperfusão de coração e pulmões pode levar a lesões celulares e até falência de órgãos no pós-operatório de cirurgia cardíaca. Vários estudos foram feitos visando melhor entendimento desses processos inflamatórios associados à circulação extracorpórea. Assim, esta revisão abordará os mecanismos de isquemia-reperfusão cardiopulmonar a ativação da cascata inflamatória por esse processo e o papel dos leucócitos na fisiopatologia destas lesões. Serão consideradas as interações de proteínas imunomoduladoras na resposta leucocitária e o bloqueio das mesmas por meio de filtros biológicos, do uso de anticorpos monoclonais e do estímulo à apoptose. / The use of extracorporeal circulation starts a whole body inflammatory response that when added with a chain reactions related to heart and lung ischemia-reperfusion may cause cellular injuries and organ failures in the cardiac surgery pos-operative period. Many studies were made to achieve better comprehension of the inflammatory process that occur with use of extracorporeal circulation. So, this review discusses the ischemia- reperfusion events, their large activated inflammatory cascade as well as the leukocytes role on the pathophysiology of their associated injuries. The interactions of immunomodulatory proteins with the leukocitary response and their blockade through leukocytes filters, the monoclonal antibody use and the apoptosis stimuli will be approached too.

Orientador: Norma Sueli Pinheiro Módolo / Banca: Leandro Gobbo Braz / Banca: Laís Helena Navarro e Lima / Banca: Rogério Luiz da Rocha Videira / Banca: Alexandra Assad / Resumo: O pós-condicionamento em modelo de isquemia e reperfusão já mostrou evidências de efeito renoprotetor, mas ainda há alguma controvérsia sobre os protocolos e seus resultados. A administração de cetamina S(+) em dose subanestésica em infusão contínua, como estratégia anti-inflamatória, ainda não foi testada na lesão renal aguda, bem como sua interação com o pós-condicionamento isquêmico não é conhecida. Testamos a hipótese de que a cetamina S(+) atenua o dano tubular e melhora a função renal em ratos sob pós-condicionamento. Quarenta e um ratos machos Wistar (≥300g) foram divididos aleatoriamente em quatro grupos: GS-Sham; GC-Cetamina S(+) em dose subanestésica em infusão contínua; GP-Pós-condicionamento isquêmico; GCP-Cetamina S(+) em dose subanestésica em infusão contínua e pós-condicionamento. Todos os animais foram submetidos à nefrectomia direita. Nos ratos submetidos ao pós-condicionamento (GP e GCP) foi realizada oclusão da artéria renal esquerda por 30 minutos. A reperfusão plena foi precedida por três ciclos de 2 min de reperfusão, seguido por 2 min de reoclusão. A pressão arterial, a frequência cardíaca e a temperatura foram controladas durante o experimento. A hidratação foi realizada com solução de Ringer lactato em infusão contínua intravenosa (3,0 mL.Kg-1.h-1), além de bolus após cada coleta. A função renal foi avaliada pela dosagem plasmática de NGAL, creatinina e ureia em três momentos: C1 (após estabilização), C2 (após 30 min de reperfusão completa) e C3 (após 24h). Dano tubular foi avaliado pela histologia renal. Foram utilizados os critérios de RIFLE e AKIN para avaliação evolutiva da creatinemia entre momentos. A creatinina e a ureia apresentaram aumento estatisticamente significativo nos grupos com pós-condicionamento isquêmico (GS e GC), mas não a NGAL (p = 0,08). Dano tubular significativo foi encontrado apenas nos ... / Abstract: Postconditioning against ischaemia-reperfusion injury has shown renoprotective effects, but there is still some controversy about protocols and its outcomes. The potencial application of subanesthetic S(+) ketamine continuous infusion as an antiinflammatory strategy, is not yet available in acute kidney injury, as well as the interaction with ischaemic postconditioning (IP). We tested the hypothesis that it attenuates tubular damage and improves renal function in IP in rats. Forty-one male Wistar rats (≥300g) were randomized into four groups: GS-sham; GK-subanesthetic S(+) ketamine; GP-posconditioning and GKP-subanesthetic S(+) ketamine and postconditioning. All animals were subjected to right nephrectomy but only in postconditioned rats 30-min left kidney arterial occlusion was performed, in which complete reperfusion were preceded by three cycles of 2 min of reperfusion followed by 2 min of reocclusion. Animals were studied for 24 h. Renal function was assessed by measurement of serum NGAL, creatinine and blood urea nitrogen (BUN) at three moments: C1 (after stabilization), C2 (after 30-min complete reperfusion) and C3 (after 24h). Tubular damage was evaluated by renal histology. RIFLE and AKIN criteria were used to evaluate creatinine among moments. Creatinine and BUN significantly increased in IP groups as compared to rats in GS and GK, but not NGAL (P=0,08). Despite significant tubular damage found only in IP groups, there was no significant difference between IP and S(+) ketamine/IP. RIFLE and AKIN criteria showed identical functional lesions. S(+) ketamine infusion does not attenuate tubular damage or improve renal function. However, IP groups show identical results and postconditioning is unable to show a renoprotective effect in this model / Doutor

Orientador: Norma Suelei Pinheiro Módolo / Banca: Geraldo Rolim Rodrigues Junior / Banca: Luiz Antonio Vane / Banca: Antonio Argolo Sampaio Filho / Banca: Rodrigo Leal Alves / Resumo: Justificativa e Objetivo: a lesão renal aguda(LRA) determina um aumento na morbidade e na mortalidade de pacientes hospitalizados. O diagnóstico precoce e a realização de medidas de proteção renal são essenciais. Foi observado que o pantoprazol possui efeito similar ao précondicionamento isquêmico em corações de ratos, promovendo efeito protetor. O objetivo deste experimento foi analisar o emprego do pantoprazol na proteção de rins submetidos à lesão de isquemia e reperfusão em ratos sob anestesia geral inalatória com isoflurano. Método: quarenta ratos, Wistar, machos, foram distribuídos aleatoriamente em quatro grupos: PIR (pantoprazol + isquemia artéria renal esquerda), P (pantoprazol), IR (isquemia renal esquerda) e Sham. O tempo de isquemia renal foi de 20 minutos e o de reperfusão, 30 minutos. Anestesia inalatória com isoflurano foi a técnica anestésica realizada. A dose administrada de pantoprazol foi de 192 mcg/kg. Todos os animais foram submetidos à nefrectomia direita e ao final do experimento procedeu-se nefrectomia esquerda, para posterior avaliação histológica e classificação por meio de escala de necrose tubular. Os atributos estudados foram: pressão arterial média (PAM), temperatura retal, os critérios de RIFLE e AKIN, dosagem urinária dos biomarcadores NGAL, KIM-1 e IL-8 e avaliação histológica. A creatinina sérica foi coletada em três momentos: M1, após monitorização; M2, após a reperfusão; e M3, 24 horas após o inicio do experimento. As coletas de urina para dosagem dos biomarcadores ocorreram antes (Urina 1) e após o experimento (Urina 2). Os atributos foram submetidos à análise estatística e as diferenças foram consideradas estatisticamente significativas quando p<0,05. Resultados: diferenças estatisticamente significativas foram observadas entre os grupos com aplicação dos critérios de RIFLE (p=0,007) e AKIN (p-0,003). Os grupos PIR e IR evoluíram com maior incidência de... / Abstract: Background and Objective: acute kidney injury (AKI) determines an increase in morbidity and mortality of hospitalized patients. Early diagnosis and the realization of renal protection measures are essential. It was observed that pantoprazole has effect similar to ischemic preconditioning in rat hearts, promoting protective effect. The objective was to analyze the use of pantoprazole in protecting kidneys subjected to ischemia and reperfusion in rats under inhalation anesthesia with isoflurane. Method: forty rats, Wistar male rats, were randomly divided into four groups: RIP (pantoprazole + left renal ischemia), P (pantoprazole), IR (left renal ischemia) and Sham. Renal ischemia time was 20 minutes and reperfusion 30 minutes. The anesthetic technique performed: inhalation anesthesia with isoflurane. The pantoprazole dose was 192 mcg/kg. All animals underwent right nephrectomy and at the end of the experiment proceeded to left nephrectomy for subsequent histological evaluation and classification through tubular necrosis scale. The parameters were: mean arterial pressure (MAP), rectal temperature, the criteria of RIFLE and AKIN, urinary dosage of biomarkers NGAL, KIM-1 and IL-8 and histological evaluation. Serum creatinine was collected in three moments: M1, after monitoring, M2, and M3 after reperfusion, 24 hours after the beginning of the experiment. The urine samples to test for biomarkers occurred before (Urine 1) and after the experiment (Urine 2). The attributes were subjected to statistical analysis and differences were considered statistically significant when p <0,05. Results: significant differences were observed between the groups with application of RIFLE criteria (p=0.007) and AKIN (p=0.003). The PIR and IR groups evolved with higher incidence of AKI. The NGAL and KIM-1 biomarkers increased in all groups throughout the experiment, but with higher values in the PIR and IR groups. IL-18 revealed no group effect (p=0.38) nor the ... / Doutor

Orientador: Winston Bonetti Yoshida / Banca: Carlos Eli Piccinato / Banca: Marcone Lima Sobreira / Banca: Ana Terezinha Guilaumon / Banca: Valter Castelli / Resumo: O uso da circulação extracorpórea desencadeia uma resposta inflamatória sistêmica, que somada às reações em cadeia relacionadas à isquemia-reperfusão de coração e pulmões pode levar a lesões celulares e até falência de órgãos no pós-operatório de cirurgia cardíaca. Vários estudos foram feitos visando melhor entendimento desses processos inflamatórios associados à circulação extracorpórea. Assim, esta revisão abordará os mecanismos de isquemia-reperfusão cardiopulmonar a ativação da cascata inflamatória por esse processo e o papel dos leucócitos na fisiopatologia destas lesões. Serão consideradas as interações de proteínas imunomoduladoras na resposta leucocitária e o bloqueio das mesmas por meio de filtros biológicos, do uso de anticorpos monoclonais e do estímulo à apoptose. / Abstract: The use of extracorporeal circulation starts a whole body inflammatory response that when added with a chain reactions related to heart and lung ischemia-reperfusion may cause cellular injuries and organ failures in the cardiac surgery pos-operative period. Many studies were made to achieve better comprehension of the inflammatory process that occur with use of extracorporeal circulation. So, this review discusses the ischemia- reperfusion events, their large activated inflammatory cascade as well as the leukocytes role on the pathophysiology of their associated injuries. The interactions of immunomodulatory proteins with the leukocitary response and their blockade through leukocytes filters, the monoclonal antibody use and the apoptosis stimuli will be approached too. / Doutor

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Objetivo: Avaliar os efeito da ornitina &#945;-cetoglutarato (OKG) no sangue e tecido intestinal de ratos submetidos à isquemia/reperfusÃo intestinal atravÃs da determinaÃÃo das concentraÃÃes in vivo no sangue e no tecido do intestino delgado, submetido a isquemia/reperfusÃo, de glicose, G 6 PDH, piruvato, acetoacetato, lactato, 3 HBDH, glutationa, T-Bars, mieloperoxidase, CPK e LDH. MÃtodo: Sessenta ratos (Rattus norvergicus albinus, Rodentia Mammalia) foram distribuÃdos aleatoriamente em cinco grupos de 12 animais: Sham 0â (s0â), Sham 30â (s30â), Sham 60â (s60â), Isquemia (i30â), ReperfusÃo (r30â). Estes grupos foram distribuÃdos em subgrupos de acordo com o tempo e com o composto utilizado na gavagem. Todos os animais receberam gavagem de caseinato de cÃlcio ou OKG em dose Ãnica, trinta minutos antes da laparotomia exploradora (LE). Os subgrupos s0âCaCa s30âCaCa, s60âc, i30âCaCa e r30âCaCa receberam apenas caseinato, de cÃlcio. Os subgrupos s0âOKG, s30âOKG, s60âOKG, i30âOKG e r30âOKG receberam OKG na dose de 5g/kg de peso. As amostras foram colhidas em cinco momentos: imediatamente apÃs a LE; apÃs 30 minutos da LE; ApÃs 1h da LE; ApÃs 30 minutos de isquemia; ApÃs 30 minutos de reperfusÃo. A estatÃstica discritiva foi expressa atravÃs da mÃdia, erro e desvio padrÃo, acompanhando-se pelo intervalo de confianÃa da mÃdia a 95% . Para comparar os valores prà e pÃs-procedimentos nas concentraÃÃes das variÃveis estudadas foram empregados os teste âtâ de Student pareado (para variÃncia homogÃnea e heterogÃnea) e ANOVA apÃs anÃlise de normalidade atravÃs do teste Kolmogorov-Smirnov. Quando observou a nÃo normalidade aplicou-se o teste de Kruskal-Wallis. Resultados: Os resultados apontaram um aumento significativo na lactacemia (1.186 + 0,18 versus 0,794 + 0,06, p<0,01) nos animais que receberam OKG em relaÃÃo ao controle nos subgrupos isquemia trinta minutos (i30â). No tecido intestinal reperfundido (r30â) ocorreu reduÃÃo significativa da concentraÃÃo de lactato (0,107 + 0,01 versus 0,266 + 0,02, p<0,05) nos animais recipientes de OKG em relaÃÃo ao animais controle, O piruvato plasmÃtico e tecidual se mostrou significantemente reduzido (0,146 + 0,24 versus 0,156 + 0,17 e 0,094 + 0,02 versus 0,248 + 0,03, p<0,05) apÃs o perÃodo de reperfusÃo de trinta minutos nos animais recipientes da OKG em relaÃÃo aos animais controle. Houve reduÃÃo significativa da concentraÃÃo do acetoacetato no tecido intestinal nos tempos pÃs isquemia e pÃs reperfusÃo dos animais recipientes da OKG (0,57 + 0,01 versus 0,0685 + 0,01 e 0,128 + 0,04 versus 0,156 + 0,03,*p<0,05) quando comparados ao animais nÃo tratados. A glicose 6 PDH apresentou reduÃÃo significativa da sua concentraÃÃo plasmÃtica no tempo isquemia trinta minutos dos animais recipientes da OKG em relaÃÃo aos nÃo tratados ( 0,1442 + 0,048 versus 1,1098 + 0,0796, *p<0,05) , ocorrendo o mesmo na concentraÃÃo tecidual, no pÃs isquemia (0,1002 + 0,02 versus 0,147 + 0,0264, p<0,05). A LDH apresentou elevaÃÃo significativa da sua concentraÃÃo nos animais recipientes da OKG em relaÃÃo ao controle (278,01 + 51,52 versus 132,93 + 12,54, *p<0,05) no grupo isquemia (i30â) . Ocorreu reduÃÃo significativa da CPK no grupo reperfusÃo (r30â) dos animais recipientes da OKG em comparaÃÃo aos animais controle (115,13 + 11,77 versus 166,70+6,23,p<0,05). A glutationa tecidual apresentou elevaÃÃo significativa no sham OKG 30 minutos em relaÃÃo ao nimais controle (59,17 + 2,39 versus 25,09 + 1.53, p<0,05) e reduÃÃo significante no tempo isquemia, tanto nos animais OKG quanto CaCa. Durante o perÃodo de reperfusÃo a glutationa nÃo apresentou alteraÃÃes significativas entre os animais tratados e controle. A OKG influenciou de maneira significativa na reduÃÃo da concentraÃÃo da 3HDBH tecidual no tempo i30â (0,062 + 0,01 versus 0,075 + 0,02,p<0,01) Esta diferenÃa significativa foi mantida no sangue dos animais tratados no grupo reperfusÃo 30âOKG em relaÃÃo aos animais r30âCaCa (0,03 + 0,00 versus 0,0615 + 0,01, p<0,01). A T-bars tecidual apresentou reduÃÃo significante no grupo r30âOKG em comparaÃÃo ao r30âCaCa (0,0522 + 0,03 versus 0,0745 + 0,02, p<0,05), com elevaÃÃo significativa no grupo sham 60âCaCa em relaÃÃo aos animais tratados 0,0937 + 0,02 versus 0,020 + 0,01, p<0,01). A oferta exÃgena da alfa-cetoglutarato nÃo ocasionou nenhuma alteraÃÃo significante nas concentraÃÃes de glicose e mieloperoxidase (MPO) entre os animais do subgrupo experimento quando comparados aos do subgrupo controle. ConclusÃes: Os procedimentos realizados foram suficientes para desencadear alteraÃÃes significativas em alguns metabÃlitos estudados. O modelo de isquemia-reperfusÃo demonstrou efetividade. A oferta exÃgena OKG, em dose Ãnica por gavagem, sugere aumento na atividade prÃ-glicolÃticas aerÃbica a nÃvel tecidual e sistÃmico; proteÃÃo contra lesÃo celular do tecido intestinal, e efeito antioxidante tecidual e sistÃmico durante a lesÃo de isquemia e apÃs a lesÃo de reperfusÃo / Objective: To evaluate the effect of ornithine alpha-ketoglutarate (OKG) in the blood and intestinal tissue of rats submitted to intestinal ischemia/reperfusion, using the blood concentrations of glucose, G6PDH, pyruvate, acetoacetate, lactate, 3HBDH, glutathione, T-Bars, myeloperoxydase, CPK and LHD, evaluated in vivo on these tissues. Methods: Sixty rats (Rattus norvergicus albinus, Rodentia Mammalia) were selected and aleatorily distributed in five groups of twelve animals, which were: Sham0â(s0â), Sham30â(s30â), Sham60â(s60â), Isquemia(i30â), ReperfusÃo(r30â). These groups were distributed in subgroups according to the time and the compost used to the âgavagemâ. All de animals received the âgavagemâ with calcium caseinate or okg, only one dosage, thirty minutes before the exploratory laparotomy (EL). The subgroups s0âCa, s30âCa, s60âCa, i30âCa and r30âCa received only calcium caseinate. The subgroups s0âokg, s30âokg, s60âokg, i30âokg and r30âokg received 5g of okg par each kilogram. The samples were taken in five moments: immediately passed the EL; passed 30 minutes of the EL; passed 60 minutes of the EL; passed 30 minutes of the isquemia; passed 30 minutes of the reperfusion. The descriptive statistics were media, error and standard deviation. The values before and after the procedures were compared using the âtâ test (âStudent pareadoâ to homogeny and heterogeny variation) and ANOVA. Then, it was used Kolmogorov-Smirnov to compare the normal results. The results were not normal, it was used the Kruskal-Wallis test. Results: It was shown an improvement on the blood lactate(1.186 + 0,18 versus 0,794 + 0,06, p<0,01) to the animals that received okg from i30â. A reduction on blood lactate lactato (0,107 + 0,01 versus 0,266 + 0,02, p<0,05) was noticed in the group r30â that received okg. It occurred a reduction on plasmatic and tissue pyruvate reduzido (0,146 + 0,24 versus 0,156 + 0,17 and 0,094 + 0,02 versus 0,248 + 0,03, p<0,05) to the group r30â that received okg. The acetoacetate was reduced to both groups, isquemia and reperfusion, that received okg0,57 + 0,01 versus 0,0685 + 0,01 e 0,128 + 0,04 versus 0,156 + 0,03,*p<0,05). The plasmatic glucose was reduced to the group i30â( 0,1442 + 0,048 versus 1,1098 + 0,0796, *p<0,05) treated with okg and the same happened to the tissue glucose after isquemia (0,1002 + 0,02 versus 0,147 + 0,0264, p<0,05). The LDH had an improvement (0,1002 + 0,02 versus 0,147 + 0,0264, p<0,05) to the group i30â treated with okg. CPK was reduced (115,13 + 11,77 versus 166,70+6,23,p<0,05) to the group r30â treated with okg. The tissue glutathione had an improvement to sham okg 30â (59,17 + 2,39 versus 25,09 + 1.53, p<0,05) and a reduction on isquemia period to the animals treated with okg and CaCa . 3HBDH was reduced (0,062 + 0,01 versus 0,075 + 0,02,p<0,01) in the blood and in the tissues from i30â. This difference was kept to animalsâ blood from r30âokg when related to r30âCaCa(0,03 + 0,00 versus 0,0615 + 0,01, p<0,01). There was a reduction on tissue T-BARS to r30âOKG when compared to r30âCaCa(0,0522 + 0,03 versus 0,0745 + 0,02, p<0,05) and an improvement to sham60âCaCa(0,0937 + 0,02 versus 0,020 + 0,01, p<0,01). Glicose and Myeloperoxydase were not affected by the use of okg. All the results were compared to the respective control groups. Conclusion: The used procedures could bring useful results to metabolites in study. The isquemia/reperfusion showed efficiency, offering exogen okg leads to a rising on glicolitic and aerobic activity to tissues and systems. This offer protects yet from the tissue injury and has antioxidant effect during the isquemia/reperfusion injury.

Activation of sympathetic nervous system is a key component of myocardial remodeling that generally occurs following ischemia/reperfusion (I/R) injury and myocardial infarction. It induces cardiac myocyte apoptosis and myocardial fibrosis, leading to myocardial dysfunction. Intracellular ubiquitin (UB) regulates protein turnover by the UB-proteosome pathway. The biological functions of extracellular UB in the heart remain largely unexplored. Previously, our lab has shown that β-adrenergic receptor (β-AR) stimulation increases extracellular UB levels, and extracellular UB inhibits β-AR-stimulated apoptosis in adult rat ventricular myocytes (ARVMs). This study explores the role of extracellular UB in myocyte apoptosis, fibroblast phenotype and function, and myocardial remodeling following β-AR stimulation and I/R injury. First, left ventricular (LV) structural and functional remodeling was studied 7 days after chronic β-AR-stimulation in the presence or absence of UB infusion. Echocardiographic analyses showed UB infusion decreases β-AR-stimulated increases in percent fractional shortening and ejection fraction. It decreased cardiac myocyte apoptosis and myocardial fibrosis. UB activated Akt, and inhibition of Akt inhibited β-AR-stimulated increases in matrix metalloproteinase-2 expression. Second, using cardiac fibroblasts, we provide evidence that extracellular UB interacts with the cell surface and co-immunoprecipitates with CXCR4. UB treatment increased expression of α-smooth muscle actin (myofibroblast marker), and induced rearrangement of actin into stress fibers. It inhibited lamellopodia and filopodia formation, and cell migration into the wound. Third, using isolated mouse heart and I/R injury as a model, we provide evidence that UB treatment decreases I/R-mediated increases in infarct size. UB treatment improved functional recovery of the heart as measured by increased % LV developed pressure. Activation of proapoptotic proteins, p-STAT-1 and caspase-9, was significantly lower in UB I/R hearts versus I/R alone. In ARVMs, UB treatment decreased simulated I/R-induced apoptosis. It activated Akt (anti-apoptotic kinase) and inhibited activation of GSK-3β (pro-apoptotic kinase). It decreased I/R-induced oxidative stress and protected anoxia-induced mitochondrial polarization. In fibroblast and ARVMs, CXCR4 antagonism negated the effects of UB, while mutated UBs (unable to interact with CXCR4) had no effect. Thus, extracellular UB, most likely acting via CXCR4, modulates myocardial remodeling with effects on heart function, fibroblast phenotype and function and myocyte apoptosis.

β-Adrenergic receptor (β-AR) stimulation increases extracellular levels of ubiquitin (UB) in myocytes, and exogenous UB decreases β-AR-stimulated myocyte apoptosis and myocardial fibrosis. Here, we hypothesized that exogenous UB modulates the inflammatory response, thereby playing a protective role in cardiac remodeling after ischemia-reperfusion (I/R) injury. C57BL/6 mice infused with vehicle or UB (1 μg·g−1·h−1) were subjected to myocardial I/R injury. Functional and biochemical parameters of the heart were examined 3 days post-I/R. Heart weight-to-body weight ratios were similarly increased in I/R and UB + I/R groups. The area at risk and infarct size were significantly lower in UB + I/R versus I/R groups. Measurement of heart function using echocardiography revealed that I/R decreases percent fractional shortening and percent ejection fraction. However, the decrease in fractional shortening and ejection fraction was significantly lower in the UB + I/R group. The UB + I/R group displayed a significant decrease in inflammatory infiltrates, neutrophils, and macrophages versus the I/R group. Neutrophil activity was significantly lower in the UB + I/R group. Analysis of the concentration of a panel of 23 cytokines/chemokines in the serum using a Bio-Plex assay revealed a significantly lower concentration of IL-12 subunit p40 in the UB + I/R versus I/R group. The concentration of monocyte chemotactic protein-1 was lower, whereas the concentration of macrophage inflammatory protein-1α was significantly higher, in the UB+I/R group versus the sham group. Expression of matrix metalloproteinase (MMP)-2 and activity of MMP-9 were higher in the UB + I/R group versus the I/R group. Levels of ubiquitinated proteins and tissue inhibitor of metalloproteinase 2 expression were increased to a similar extent in both I/R groups. Thus, exogenous UB plays a protective role in myocardial remodeling post-I/R with effects on cardiac function, area at risk/infarct size, the inflammatory response, levels of serum cytokines/chemokines, and MMP expression and activity.