京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18645号 / 医博第3944号 / 新制||医||1006(附属図書館) / 31559 / 京都大学大学院医学研究科医学専攻 / (主査)教授 伊達 洋至, 教授 羽賀 博典, 教授 千葉 勉 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20081号 / 医博第4174号 / 新制||医||1018(附属図書館) / 33197 / 京都大学大学院医学研究科医学専攻 / (主査)教授 湊谷 謙司, 教授 福田 和彦, 教授 浅野 雅秀 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Objective: The aim of this work is to evaluate the effect of warm ischemia on renal function. Methods: Within a multicentric study, the data about tumor-affected solitary kidney were obtained from 9 urological centers in the Czech Republic. Patients were divided into groups according to the WIT (warm ischemia time). In each group the mean preoperative serum creatinine was determined, as well as on 3rd and 7th postoperative day and the lowest GF postoperatively. In each group the mean R.E.N.A.L. nephrometric score was determined. Results: The study compriese data totally of 97 patients. The open approach was chosen in 78 cases, in 16 cases laparoscopic approach was chosen. The robotic surgery was performed in 3 patients. PN with zero ischemia was performed in 29 patients (30%). Conclusion: Our findings confirm that non-clamping partial nephrectomy in a solitary kidney minimizes renal injury. For the non-clamping partial nephrectomy can be mainly indicated smaller exophyticall tumors without deep parenchymal invasion. Unfortunately, most of PN should be performed with vascular clamping, allowing precise closure of collecting system, vascular and parenchymal defect, especially for larger tumors with deep parenchymal invasion. According to our observation that WIT of 15 minutes for a kidney is safe. Clamping of...

Objective: The aim of this work is to evaluate the effect of warm ischemia on renal function. Methods: Within a multicentric study, the data about tumor-affected solitary kidney were obtained from 9 urological centers in the Czech Republic. Patients were divided into groups according to the WIT (warm ischemia time). In each group the mean preoperative serum creatinine was determined, as well as on 3rd and 7th postoperative day and the lowest GF postoperatively. In each group the mean R.E.N.A.L. nephrometric score was determined. Results: The study compriese data totally of 97 patients. The open approach was chosen in 78 cases, in 16 cases laparoscopic approach was chosen. The robotic surgery was performed in 3 patients. PN with zero ischemia was performed in 29 patients (30%). Conclusion: Our findings confirm that non-clamping partial nephrectomy in a solitary kidney minimizes renal injury. For the non-clamping partial nephrectomy can be mainly indicated smaller exophyticall tumors without deep parenchymal invasion. Unfortunately, most of PN should be performed with vascular clamping, allowing precise closure of collecting system, vascular and parenchymal defect, especially for larger tumors with deep parenchymal invasion. According to our observation that WIT of 15 minutes for a kidney is safe. Clamping of...

INTRODUÇÃO: O limite de tempo de isquemia renal sem causar um dano permanente à função ainda é um questionamento pertinente na prática cirúrgica urológica. Algumas cirurgias requerem um campo cirúrgico menos sangrante, necessitando de uma interrupção temporária do fluxo sanguíneo para o parênquima renal através de clampeamento do pedículo. Este estudo objetivou avaliar qual técnica de clampeamento em isquemia isotérmica é superior, em termos de preservação da função renal. MÉTODOS: Neste estudo, foram utilizados 28 coelhos da raça New Zealand distribuídos em quatro grupos de forma randomizada e submetidos à laparotomia com interrupção do fluxo sanguíneo renal esquerdo, conforme grupo: grupo 1 - Controle, sem isquemia (4 animais), grupo 2 - Isquemia seletiva (8 animais), grupo 3 - Isquemia não-seletiva (8 animais) e grupo 4 - Isquemia não-seletiva intermitente (8 animais). Os animais foram submetidos à análises cintilográficas da função renal com mercaptoacetiltriglicina (MAG3) e dosagens séricas de creatinina no pré-operatório, no primeiro, terceiro e sétimo dias pós-operatórios. Após 15 dias da cirurgia, os animais foram sacrificados e os rins submetidos à análise histológica. RESULTADOS: A análise cintilográfica da função renal mostrou que os três grupos submetidos à isquemia apresentaram um agravo semelhante, nas primeiras 24 horas; não havendo diferença estatística entre eles no tocante a perda de função renal do rim esquerdo (p= 0,165), neste período. No exame do terceiro dia pós-operatório, houve diferença estatística (p= 0,006) entre os grupos não-seletivo (3) e o intermitente (4), mostrando uma superioridade do fator desclampeamento na proteção à função renal. No sétimo dia pós-operatório, o grupo do clampeamento seletivo (2) mostrou-se superior (p< 0,001) ao grupo não-seletivo (3), mas não apresentou diferença estatística em relação ao grupo intermitente (4). Este último, por sua vez, também apresentou-se superior ao grupo 3, com diferença estatística (p< 0,001). Após sete dias não foi observada diferença estatística entre os grupos 2 e 4. A histologia e a creatinina não mostraram diferença estatisticamente significante entre os grupos isquêmicos. Do ponto de vista funcional, a isquemia seletiva e a intermitente foram superiores em relação a não-seletiva. CONCLUSÃO: O modelo utilizado neste estudo mostrou uma superioridade da isquemia isotérmica seletiva e da intermitente em relação a não-seletiva na proteção da função renal. Não houve diferença entre o clampeamento seletivo e o intermitente não-seletivo na função do rim / INTRODUCTION: The time limit of renal ischemia with no permanent damage to renal function still remains as a pertinent question in urologic surgical practice. Several procedures require a bloodless surgical field most of time needing an interruption of blood flow to the renal parenchyma and they can require hilum control and clamping. This study assessed which normothermic clamping technique is superior in order to preserve kidney function. METHODS: In this project, 28 New Zealand rabbits were distributed in four groups by randomization and submitted to laparotomy and blood flow interruption of left kidney: Group 1 - Control, no ischemia (four animals), Group 2 Selective ischemia (eight animals), Group 3 Nonselective ischemia (eight animals) e Group 4 intermittent nonselective ischemia (eight animals). The animals were submitted to renal scintigraphy with mercaptoacetiltriglicin (MAG3) and serum creatinine analysis preoperatively and at days 1, 3 and 7 after left hilum clamping. The animals were submitted to nephrectomy and immediately sacrificed and the kidneys subjected to blinded evaluation by a nephropathologist. RESULTS: Renal scintigraphy showed equal damage in the three groups in first 24 hours, with no statistical difference in loss of left kidney function (p= 0,165) in this period. At third day examination, there was statistical difference (p= 0,006) between the nonselective group (3) and intermittent one (4), demonstrating a superiority of declamping factor in renal function protection. At seventh postoperative day, selective group (2) proved to be better (p< 0,001) than nonselective one (3), however there was not statistical difference between group 2 and the intermittent group (4). The intermittent clamping group was also superior to group 3 (p< 0,001). Histopathology and serum creatinine did not demonstrate statistical difference among groups. Functionally, selective and intermittent warm ischemia techniques are better than nonselective one. CONCLUSION: The model used in this study presented a superiority of selective clamping and intermittent arteriovenous clamping in shielding the renal function. No difference occurred between selective clamping and intermittent nonselective one

A trombose venosa é a principal complicação da microcirurgia vascular e a intervenção precoce é necessária para o salvamento dos retalhos, com índices de sucesso de apenas 50% das revisões cirúrgicas; trombose da microcirculação, produção de radicais livres de oxigênio (RLO) e edema são os elementos principais da lesão de isquemia/reperfusão (I/R), e o planejamento das terapias protetoras tem como objetivo amenizar estas alterações. Os fármacos antioxidantes, antiagregantes plaquetários e anticoagulantes são utilizados no controle da lesão de I/R em diferentes órgãos. Neste estudo, em modelo de amputação subtotal de membro posterior de rato submetido a isquemia global primária, foi testado o efeito protetor dos fármacos alopurinol, heparina, tirofiban ou vitamina C durante a isquemia secundaria pós congestão venosa. Foram operados 100 ratos, que apos isquemia global de 90 minutos, foram divididos em cinco grupos de 20 animais recebendo uma das respectivas drogas na veia femoral contra-lateral: 1ml de solução fisiológica 0,9% no grupo controle (GC), 1ml de alopurinol 45mg/kg no grupo experimental 1 (G1), 1ml de heparina 200UI/kg no grupo experimental 2 (G2), 1ml de tirofiban 50 ug /ml no grupo experimental 3 (G3) e 1 ml de vitamina C 100mg/kg no grupo experimental 4 (G4); o clampe foi então retirado do feixe vascular e se iniciou a reperfusão de 60 minutos; a colocação do clampe vascular apenas na veia femoral direita iniciou a congestão venosa (isquemia secundária) do membro por 90 minutos seguido de outra reperfusão de 60 minutos; O músculo gastrocnêmio foi dissecado e retirado para analise histológica e os animais sacrificados por injeção letal. Foram estudados a porcentagem de viabilidade celular muscular, o edema e o extravasamento de hemácias. A porcentagem de lesão celular do músculo do grupo controle foi 54,6% (±10,6), do G1 31,5% (±13,6), do G2 24,7% (±11,7), do G3 24,6% (±8,6) e do G4 21,3% (±8,6). Os grupos foram comparados por modelo de comparação múltiplas one way-ANOVA e post-hoc Tukey com significância de p < 0,05. A porcentagem de lesão celular foi menor para os grupos G1, G2, G3 e G4 quando comparados ao GC (p < 0,001), e quando comparados os grupos experimentais entre si, apenas o G4 (vitamina C) foi menor estatisticamente que G1(alopurinol) (p < 0,029). A utilização individual dos fármacos alopurinol, heparina ,tirofiban e vitamina C mostraram efeito protetor na congestão venosa secundaria a isquemia global primária, e a vitamina C foi mais efetiva nesta ação que o alopurinol quando comparados os antioxidantes entre si. Quando avaliado o edema, apenas os antioxidantes tiveram índices menores estatisticamente que o GC, enquanto que todos os fármacos diminuíram o extravasamento de hemácias comparados com o grupo controle (p < 0,001) / Venous thrombosis is the main complication of vascular microsurgery an early intervention is mandatory to rescue the flap, with a success rate of only 50% of surgical revisions; microcirculation thrombosis, oxygen free radicals production and edema are the main elements of ischemia/reperfusion (I/R) injury, and protective therapies aim to mitigate these changes. Antioxidants, antiplatelets and anticoagulants are used in different organs to control this injury. In this study, in a partial hind limb amputation model submitted to global ischemia, it was tested the protective effect of Allopurinol, Heparin, Tirofiban or Vitamin C during secondary ischemia after venous congestion. A hundred rats divided in five groups of 20 animals each were operated; after global ischemia of 90 minutes each group was injected into the contra lateral femoral vein one of the following solutions: 1 ml of saline solution NaCl 0,9% - control group (CG); 1ml of Allopurinol 45mg/kg - experimental group 1 (G1); 1ml of Heparin 200 UI/kg - experimental group 2 (G2); 1ml of Tirofiban 50 ug /ml - experimental group 3 (G3); 1ml of Vitamin C 100mg/kg - experimental group 4 (G4). Sixty minutes of limb reperfusion was performed, and a secondary period of limb ischemia started with the clamping of the femoral vein only (limb congestion) which lasted for 90 minutes (secondary ischemia). After that, the vein clamp was removed and a 60 minute reperfusion period was observed; at the end of the second reperfusion period, the right gastrocnemius muscle was removed and fixed in 10% formaldehyde, animals were euthanized with a lethal dose of Pentobarbital. Muscle fibers were scored as uninjured or injured based on the morphology of individual fibers; interstitial edema and bleeding were graded on a four-point scale. The control group had more damaged muscle cells 54.6±10.6% when compared to allopurinol 31.5±13,6%, heparin 24.7±11.7%, tirofiban 24.6±8.6% and Vitamin C 21.3±8.6% all reached statistical significance (p < 0.00 0.029). These comparisons were analysed using ANOVA and post-hoc Tukey. The single use of Allopurinol, Heparin, Tirofiban or Vitamin C showed a protective effect on venous congestion after global ischemia, and Vitamin C was more effective than Allopurinol when compared both antioxidants. When evaluating the edema, only the antioxidants had statistically lower rates than the CG, whilst all drugs reduced the extravasation of red blood cells compared with the control group (p < 0.001)

INTRODUÇÃO: O limite de tempo de isquemia renal sem causar um dano permanente à função ainda é um questionamento pertinente na prática cirúrgica urológica. Algumas cirurgias requerem um campo cirúrgico menos sangrante, necessitando de uma interrupção temporária do fluxo sanguíneo para o parênquima renal através de clampeamento do pedículo. Este estudo objetivou avaliar qual técnica de clampeamento em isquemia isotérmica é superior, em termos de preservação da função renal. MÉTODOS: Neste estudo, foram utilizados 28 coelhos da raça New Zealand distribuídos em quatro grupos de forma randomizada e submetidos à laparotomia com interrupção do fluxo sanguíneo renal esquerdo, conforme grupo: grupo 1 - Controle, sem isquemia (4 animais), grupo 2 - Isquemia seletiva (8 animais), grupo 3 - Isquemia não-seletiva (8 animais) e grupo 4 - Isquemia não-seletiva intermitente (8 animais). Os animais foram submetidos à análises cintilográficas da função renal com mercaptoacetiltriglicina (MAG3) e dosagens séricas de creatinina no pré-operatório, no primeiro, terceiro e sétimo dias pós-operatórios. Após 15 dias da cirurgia, os animais foram sacrificados e os rins submetidos à análise histológica. RESULTADOS: A análise cintilográfica da função renal mostrou que os três grupos submetidos à isquemia apresentaram um agravo semelhante, nas primeiras 24 horas; não havendo diferença estatística entre eles no tocante a perda de função renal do rim esquerdo (p= 0,165), neste período. No exame do terceiro dia pós-operatório, houve diferença estatística (p= 0,006) entre os grupos não-seletivo (3) e o intermitente (4), mostrando uma superioridade do fator desclampeamento na proteção à função renal. No sétimo dia pós-operatório, o grupo do clampeamento seletivo (2) mostrou-se superior (p< 0,001) ao grupo não-seletivo (3), mas não apresentou diferença estatística em relação ao grupo intermitente (4). Este último, por sua vez, também apresentou-se superior ao grupo 3, com diferença estatística (p< 0,001). Após sete dias não foi observada diferença estatística entre os grupos 2 e 4. A histologia e a creatinina não mostraram diferença estatisticamente significante entre os grupos isquêmicos. Do ponto de vista funcional, a isquemia seletiva e a intermitente foram superiores em relação a não-seletiva. CONCLUSÃO: O modelo utilizado neste estudo mostrou uma superioridade da isquemia isotérmica seletiva e da intermitente em relação a não-seletiva na proteção da função renal. Não houve diferença entre o clampeamento seletivo e o intermitente não-seletivo na função do rim / INTRODUCTION: The time limit of renal ischemia with no permanent damage to renal function still remains as a pertinent question in urologic surgical practice. Several procedures require a bloodless surgical field most of time needing an interruption of blood flow to the renal parenchyma and they can require hilum control and clamping. This study assessed which normothermic clamping technique is superior in order to preserve kidney function. METHODS: In this project, 28 New Zealand rabbits were distributed in four groups by randomization and submitted to laparotomy and blood flow interruption of left kidney: Group 1 - Control, no ischemia (four animals), Group 2 Selective ischemia (eight animals), Group 3 Nonselective ischemia (eight animals) e Group 4 intermittent nonselective ischemia (eight animals). The animals were submitted to renal scintigraphy with mercaptoacetiltriglicin (MAG3) and serum creatinine analysis preoperatively and at days 1, 3 and 7 after left hilum clamping. The animals were submitted to nephrectomy and immediately sacrificed and the kidneys subjected to blinded evaluation by a nephropathologist. RESULTS: Renal scintigraphy showed equal damage in the three groups in first 24 hours, with no statistical difference in loss of left kidney function (p= 0,165) in this period. At third day examination, there was statistical difference (p= 0,006) between the nonselective group (3) and intermittent one (4), demonstrating a superiority of declamping factor in renal function protection. At seventh postoperative day, selective group (2) proved to be better (p< 0,001) than nonselective one (3), however there was not statistical difference between group 2 and the intermittent group (4). The intermittent clamping group was also superior to group 3 (p< 0,001). Histopathology and serum creatinine did not demonstrate statistical difference among groups. Functionally, selective and intermittent warm ischemia techniques are better than nonselective one. CONCLUSION: The model used in this study presented a superiority of selective clamping and intermittent arteriovenous clamping in shielding the renal function. No difference occurred between selective clamping and intermittent nonselective one

A trombose venosa é a principal complicação da microcirurgia vascular e a intervenção precoce é necessária para o salvamento dos retalhos, com índices de sucesso de apenas 50% das revisões cirúrgicas; trombose da microcirculação, produção de radicais livres de oxigênio (RLO) e edema são os elementos principais da lesão de isquemia/reperfusão (I/R), e o planejamento das terapias protetoras tem como objetivo amenizar estas alterações. Os fármacos antioxidantes, antiagregantes plaquetários e anticoagulantes são utilizados no controle da lesão de I/R em diferentes órgãos. Neste estudo, em modelo de amputação subtotal de membro posterior de rato submetido a isquemia global primária, foi testado o efeito protetor dos fármacos alopurinol, heparina, tirofiban ou vitamina C durante a isquemia secundaria pós congestão venosa. Foram operados 100 ratos, que apos isquemia global de 90 minutos, foram divididos em cinco grupos de 20 animais recebendo uma das respectivas drogas na veia femoral contra-lateral: 1ml de solução fisiológica 0,9% no grupo controle (GC), 1ml de alopurinol 45mg/kg no grupo experimental 1 (G1), 1ml de heparina 200UI/kg no grupo experimental 2 (G2), 1ml de tirofiban 50 ug /ml no grupo experimental 3 (G3) e 1 ml de vitamina C 100mg/kg no grupo experimental 4 (G4); o clampe foi então retirado do feixe vascular e se iniciou a reperfusão de 60 minutos; a colocação do clampe vascular apenas na veia femoral direita iniciou a congestão venosa (isquemia secundária) do membro por 90 minutos seguido de outra reperfusão de 60 minutos; O músculo gastrocnêmio foi dissecado e retirado para analise histológica e os animais sacrificados por injeção letal. Foram estudados a porcentagem de viabilidade celular muscular, o edema e o extravasamento de hemácias. A porcentagem de lesão celular do músculo do grupo controle foi 54,6% (±10,6), do G1 31,5% (±13,6), do G2 24,7% (±11,7), do G3 24,6% (±8,6) e do G4 21,3% (±8,6). Os grupos foram comparados por modelo de comparação múltiplas one way-ANOVA e post-hoc Tukey com significância de p < 0,05. A porcentagem de lesão celular foi menor para os grupos G1, G2, G3 e G4 quando comparados ao GC (p < 0,001), e quando comparados os grupos experimentais entre si, apenas o G4 (vitamina C) foi menor estatisticamente que G1(alopurinol) (p < 0,029). A utilização individual dos fármacos alopurinol, heparina ,tirofiban e vitamina C mostraram efeito protetor na congestão venosa secundaria a isquemia global primária, e a vitamina C foi mais efetiva nesta ação que o alopurinol quando comparados os antioxidantes entre si. Quando avaliado o edema, apenas os antioxidantes tiveram índices menores estatisticamente que o GC, enquanto que todos os fármacos diminuíram o extravasamento de hemácias comparados com o grupo controle (p < 0,001) / Venous thrombosis is the main complication of vascular microsurgery an early intervention is mandatory to rescue the flap, with a success rate of only 50% of surgical revisions; microcirculation thrombosis, oxygen free radicals production and edema are the main elements of ischemia/reperfusion (I/R) injury, and protective therapies aim to mitigate these changes. Antioxidants, antiplatelets and anticoagulants are used in different organs to control this injury. In this study, in a partial hind limb amputation model submitted to global ischemia, it was tested the protective effect of Allopurinol, Heparin, Tirofiban or Vitamin C during secondary ischemia after venous congestion. A hundred rats divided in five groups of 20 animals each were operated; after global ischemia of 90 minutes each group was injected into the contra lateral femoral vein one of the following solutions: 1 ml of saline solution NaCl 0,9% - control group (CG); 1ml of Allopurinol 45mg/kg - experimental group 1 (G1); 1ml of Heparin 200 UI/kg - experimental group 2 (G2); 1ml of Tirofiban 50 ug /ml - experimental group 3 (G3); 1ml of Vitamin C 100mg/kg - experimental group 4 (G4). Sixty minutes of limb reperfusion was performed, and a secondary period of limb ischemia started with the clamping of the femoral vein only (limb congestion) which lasted for 90 minutes (secondary ischemia). After that, the vein clamp was removed and a 60 minute reperfusion period was observed; at the end of the second reperfusion period, the right gastrocnemius muscle was removed and fixed in 10% formaldehyde, animals were euthanized with a lethal dose of Pentobarbital. Muscle fibers were scored as uninjured or injured based on the morphology of individual fibers; interstitial edema and bleeding were graded on a four-point scale. The control group had more damaged muscle cells 54.6±10.6% when compared to allopurinol 31.5±13,6%, heparin 24.7±11.7%, tirofiban 24.6±8.6% and Vitamin C 21.3±8.6% all reached statistical significance (p < 0.00 0.029). These comparisons were analysed using ANOVA and post-hoc Tukey. The single use of Allopurinol, Heparin, Tirofiban or Vitamin C showed a protective effect on venous congestion after global ischemia, and Vitamin C was more effective than Allopurinol when compared both antioxidants. When evaluating the edema, only the antioxidants had statistically lower rates than the CG, whilst all drugs reduced the extravasation of red blood cells compared with the control group (p < 0.001)

Aim: Complex arterial reconstruction in kidney transplantation (KT) using kidneys from deceased donors (DD) warrants additional study since little is known about the effects on the mid- and long-term outcome and graft survival. Methods: A total of 451 patients receiving deceased donor KT in our department between 1993 and 2017 were included in our study. Patients were divided into three groups according to the number of arteries and anastomosis: (A) 1 renal artery, 1 arterial anastomosis (N = 369); (B) >1 renal artery, 1 arterial anastomosis (N = 47); and (C) >1 renal artery, >1 arterial anastomosis (N = 35). Furthermore, the influence of localization of the arterial anastomosis (common iliac artery (CIA), versus non-CIA) was analyzed. Clinicopathological characteristics, outcome, and graft and patient survival of all groups were compared retrospectively. Results: With growing vascular complexity, the time of warm ischemia increased significantly (groups A, B, and C: 40 ± 19 min, 45 ± 19 min, and 50 ± 17 min, respectively; p = 0.006). Furthermore, the duration of operation was prolonged, although this did not reach significance (groups A, B, and C: 175 ± 98 min, 180 ± 35 min, and 210 ± 43 min, respectively; p = 0.352). There were no significant differences regarding surgical complications, post-transplant kidney function (delayed graft function, initial non-function, episodes of acute rejection), or long-term graft survival. Regarding the localization of the arterial anastomosis, non-CIA was an independent prognostic factor for deep vein thrombosis in multivariate analysis (CIA versus non-CIA: OR 11.551; 95% CI, 1.218–109.554; p = 0.033). Conclusion: Multiple-donor renal arteries should not be considered a contraindication to deceased KT, as morbidity rates and long-term outcomes seem to be comparable with grafts with single arteries and less complex anastomoses.

Contexte : Au cours de la dernière décennie, le nombre de personnes en attente d’une transplantation cardiaque a augmenté d’environ 25%, tandis que le nombre de greffes effectuées chaque année est resté stable. Le taux de décès des patients en attente d’une greffe cardiaque est d’environ 15-20%. Le don d’organe suite à un décès cardiocirculatoire (DDC) est une alternative au don après décès neurologique (DDN) qui a permis d’augmenter le nombre d’organes disponibles comme les poumons, les reins et les foies. Compte tenu de la survenue d’une mort cardiovasculaire dans les protocoles DDC, le cœur est rarement greffé à cause des dommages infligés durant la période d’ischémie chaude. Notre équipe a précédemment démontré que l’utilisation du Celastrol, ainsi que notre analogue synthétique inhibiteur de la HSP90 ont des effets cardioprotecteurs, quand administrés au moment de la reperfusion dans des modèles in vitro de culture cellulaire et ex vivo dans des cœurs de rats montés sur le système de perfusion Langendorff. L’objectif est d’évaluer les mécanismes cardioprotecteurs rapides d’une nouvelle formulation de l’inhibiteur HSP90, et de comprendre l’efficacité de ce nouveau composé synthétique sur deux lignées de cellules : les cardiomyoblastes H9c2 issus de rats et les cardiomyocytes dérivés de cellules souches pluripotentes humaines (iPSC-CMs). Méthodes/Résultats : Les cellules H9c2 et iPSC-CMs ont été cultivées. La signalisation cellulaire a été analysée par western blot pour évaluer le niveau d’activation de ces différentes voies. Suite à l’optimisation des conditions pour les cellules iPSC-CMs, les deux lignées cellulaires ont été mises en condition ischémique (sans glucose, 95% N2, 5% CO2) durant la nuit, puis reperfusées, en conditions normales, avec différentes concentrations de l’inhibiteur HSP90. La viabilité cellulaire ainsi que l’ouverture des pores mitochondriaux (mPTP) ont été évaluées à l’aide de kits d’analyses, la production de radicaux libres d’oxygène à l’aide de kits de fluorescence et l’expression des ARN messagers de gènes antioxydants à l’aide de la réaction en chaîne par polymérase (PCR). Les résultats ont montré une augmentation de l’activation des voies cytoprotectrices quand les deux lignées cellulaires étaient traitées à la concentration 10-6M du composé sans stress 4 ischémique : augmentation de HO-1 and HSP-70 dans les 30 premières minutes et AKT et ERK après 1 heure de traitement et 3 heures de récupération. Contrairement à nos attentes, le traitement au moment de la reperfusion à la concentration 10-6M a montré une diminution de la viabilité des cellules, alors que la concentration 10-7M l’a augmenté. À une concentration de 10- 7M, il y a eu diminution de la production de radicaux libres comparativement au groupe témoin. Comme attendu, cette concentration a aussi démontré une diminution de l’ouverture des mPTP. Tous ces résultats ont été observés, autant dans les cellules humaines que celles de rats. Une évaluation préliminaire de l’expression des gènes antioxydants dans les cellules H9c2 a seulement montré une augmentation de l’expression des gènes CAT et HO-1. Conclusion : Notre groupe de recherche a précédemment démontré l’efficacité des composés issus du Celastrol sur la réduction des dommages myocardiques dus à la reperfusion dans les modèles d’ischémie, incluant l’infarctus du myocarde et la donation après décès cardiocirculatoire. Ces expériences ont montré les effets bénéfiques du nouveau composé synthétique sur l’expression des gènes antioxydants, et sur l’activation d’une série de voies cytoprotectrices permettant la stabilisation de la membrane mitochondriale, réduisant aussi la production de radicaux libres, et améliorant ultimement la survie cellulaire. Des études supplémentaires sont en cours afin d’améliorer la compréhension des modes d’action, des mécanismes et des dosages optimaux du médicament, ce qui nous permettra de commencer les essais sur animaux dans le but d’introduire cette molécule en clinique dans le contexte de don d’organes. / Background: During the last decade, the number of people waiting for a cardiac transplantation has increased by about 25%, while the number of yearly transplant surgeries performed has remained steady. The death rate of patients awaiting heart transplant is about 15-20%. Organ donation after circulatory death (DCD) is an alternative to donation after neurological death (DND) that has allowed to increase the number of available organs like lungs, livers, and kidneys. However, because of the cardiac death in DCD protocols, the heart is rarely used because of the injuries suffered by the warm ischemia period. Our group has previously shown that Celastrol, along with a synthetic HSP90 inhibitor analog, have cardioprotective effects when given as postconditioning agents at the moment of reperfusion in an in vitro model on cellular cultures and an ex vivo model on rat hearts mounted on a Langendorff perfusion system. The objective is to evaluate the rapid cardioprotective mechanisms of a novel formulation of the HSP90 inhibitor compound, and to understand the efficacy of this new synthetic compound on two cell lines: rat H9c2 cardiomyoblasts and human induced pluripotent stem cell-derived cardiomyocytes (iPSCCMs). Methods/Results: H9c2 rat cardiomyoblasts and human iPSC-CMs were cultured. Cell signaling was analyzed by western blot to evaluate pathway activations. Both cell lines were put in ischemic conditions (no glucose, 95% N2, 5% CO2) overnight, then reperfused (normal conditions) with different concentrations of HSP90i after optimizing the human iPSC-CMs’ stress experiment. Cell viability and mitochondrial permeability transition pore (mPTP) opening were evaluated using assays, oxygen-free radical production by fluorescence assay and antioxidant gene messenger RNA expression via polymerase chain reaction (PCR). Results showed an increase in cytoprotective pathway activation when both cell lines were treated with 10-6M of the compound without any stress: HO-1 and HSP-70 in the first 30 minutes while AKT and ERK after 1 hour of treatment and 3 hours of recuperation. Interestingly, treatment with the compound at 10-6M at the moment of reperfusion showed decreased viability of the cells while 10-7M improved it. Free radical production was also decreased at a concentration of 10-7M 6 when compared to the baseline, and as expected, the compound also decreased mPTP opening. These results were seen in both human and rat cell lines. Preliminary evaluation of antioxidant gene expression in H9c2 cells only showed an increase in the expression of the cytoprotective CAT and HO-1 genes. Conclusion: Our research group has previously demonstrated the efficacy of Celastrol compounds in reducing reperfusion damage in myocardial ischemia models, including myocardial infarction and donation after circulatory death. These experiments have shown that the beneficial effects of this new synthetic compound include the expression of antioxidant genes and the launching of a series of cytoprotective pathways that stabilize the mitochondrial membrane, reduce free radical production, and improve cell survival. Additional studies to fully understand the mode of action, the mechanisms and the optimal dosages are underway to allow us to move to animal trials in order to ultimately introduce the molecule in the clinical field in the context of organ donation.