京都大学 / 新制・課程博士 / 博士(医学) / 甲第24518号 / 医博第4960号 / 新制||医||1065(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 柳田 素子, 教授 江木 盛時, 教授 長船 健二 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Abstract There is a lack of reliable methods of evaluation of brain ischemic injury in patients undergoing cardiac surgery. The present study was, therefore, planned to evaluate whether serum S100β protein (I), brain cortical microdialysis (II), intracranial pressure (III) and electroencephalography (EEG) (IV) are predictive of postoperative death and brain ischemic injury in an experimental surviving porcine model of hypothermic circulatory arrest (HCA). One hundred and twenty eight (128) female, juvenile (8 to 10 weeks of age) pigs of native stock, weighing 21.0 to 38.2 kg, underwent cardio-pulmonary bypass prior to, and following, a 75-minute period of HCA at a brain temperature of 18°C. During the operation, hemodynamic, electrocardiograph and temperature monitoring was performed continuously. Furthermore, metabolic parameters were monitored at baseline, end of cooling, at intervals of two, four and eight hours after HCA and before extubation. Electroencephalographic recording was performed in all animals, serum S100β protein measurement in 18 animals, cortical microdialysis in 109 animals, and intracranial pressure monitoring in 58 animals. After the operation, assessment of behavior was made on a daily basis until death or elective sacrifice on the seventh postoperative day. All four studies showed that these parameters were predictive of postoperative outcome. Animals with severe histopathological injury had higher serum S100β protein levels at every time interval after HCA. Analysis of cortical brain microdialysis showed that the lactate/glucose ratio was significantly lower and the brain glucose concentration significantly higher among survivors during the early postoperative hours. Intracranial pressure increased significantly after 75 minutes of HCA, and this was associated with a significantly increased risk of postoperative death and brain infarction. A slower recovery of EEG burst percentage after HCA was significantly associated with the development of severe cerebral cortex, brain stem and cerebellum ischemic injury. In conclusion, serum S100β protein proved to be a reliable marker of brain ischemic injury as assessed on histopathological examination. Cerebral microdialysis is a useful method of cerebral monitoring during experimental HCA. Low brain glucose concentrations and high brain lactate/ glucose ratios after HCA are strong predictors of postoperative death. Increased intracranial pressure severely affected the postoperative outcome and may be a potential target for treatment. EEG burst percentage as a sum effect of anesthetic agent and ischemic brain damage is a useful tool for early prediction of severe brain damage after HCA. Among these monitoring methods, brain cortical microdialysis seems to be the most powerful one in predicting brain injury after experimental hypothermic circulatory arrest.

Devido às dificuldades encontradas na Medicina Veterinária com relação à circulação extracorpórea, despertou-se o interesse pela técnica de parada circulatória total (\"Inflow Occlusion\") para correção cirúrgica de algumas patologias cardíacas, principalmente estenose de valvas semi-lunares. O objetivo deste trabalho consistiu em comparar as possíveis alterações hemodinâmicas que podem ocorrer em cães submetidos à parada circulatória total através da técnica do \"Inflow Occlusion\" por diferentes períodos de tempo. Utilizou-se neste estudo, doze cães, subdivididos em dois grupos, sendo submetidos à parada circulatória total, por sete ou oito minutos. Realizou-se análise hemodinâmica durante quatro momentos do período transoperatório. Os parâmetros de oxigenação foram obtidos por meio da conjunção dos dados fornecidos pelo cateter de artéria pulmonar e pela análise hemogasométrica. Diferença estatisticamente significativa foi encontrada na comparação entre os grupos avaliados, cinco minutos após a restituição da circulação (momento M3), para a pressão arterial média (PAM) e para o índice de trabalho sistólico do ventrículo direito (ITSVD) e do ventrículo esquerdo (ITSVE). Ocorreram dois óbitos transoperatórios no grupo de animais submetidos à oito minutos de parada circulatória total, devido a fibrilação ventricular. Frente aos óbitos ocorridos, conclui-se que o \"Inflow Occlusion\" foi exequível em cães sadios por sete minutos, sendo contra-indicado para períodos de oito minutos. Entretanto, estudos clínicos envolvendo a técnica em questão, precisam ser realizados para que a referida técnica possa ser aplicada, seguramente, em animais portadores de disfunções cardiovasculares. / Because of difficulties related to extracorporeal circulation in Veterinary Medicine, more interest occurred about circulatory arrest trough \"Inflow Occlusion\" technique to correct surgically some cardiac pathologies, such as semilunar valves stenosis. The aim of this study was to compare possible hemodynamic alterations that can occur in dogs submitted to circulatory arrest using \"Inflow Occlusion\" technique for different periods of time. Twelve mongrel dogs were used in this study divided in two groups, each one submitted to seven or eight minutes of total circulatory arrest. Hemodynamic evaluation was realized during four moments on transoperatory period. Oxygenation parameters were obtained trough pulmonary artery catheter data and hemogasometric analysis. Statistical difference between groups were found on M3 moment, five minutes after circulatory arrest, for mean arterial pressure (MAP), right ventricular systolic stroke index (RVSSI) and left ventricular systolic stroke index (LVSSI). There were two transoperatory deaths on group B, because ventricular fibrillation. Because these deaths, we can say that \"Inflow Occlusion\" can be realized up to seven minutes on healthy dogs and is contra-indicated for periods of eight minutes. However, clinical studies should be performed in order to use this technique routinely with safe in dogs presenting cardiovascular abnormalities.

Devido às dificuldades encontradas na Medicina Veterinária com relação à circulação extracorpórea, despertou-se o interesse pela técnica de parada circulatória total (\"Inflow Occlusion\") para correção cirúrgica de algumas patologias cardíacas, principalmente estenose de valvas semi-lunares. O objetivo deste trabalho consistiu em comparar as possíveis alterações hemodinâmicas que podem ocorrer em cães submetidos à parada circulatória total através da técnica do \"Inflow Occlusion\" por diferentes períodos de tempo. Utilizou-se neste estudo, doze cães, subdivididos em dois grupos, sendo submetidos à parada circulatória total, por sete ou oito minutos. Realizou-se análise hemodinâmica durante quatro momentos do período transoperatório. Os parâmetros de oxigenação foram obtidos por meio da conjunção dos dados fornecidos pelo cateter de artéria pulmonar e pela análise hemogasométrica. Diferença estatisticamente significativa foi encontrada na comparação entre os grupos avaliados, cinco minutos após a restituição da circulação (momento M3), para a pressão arterial média (PAM) e para o índice de trabalho sistólico do ventrículo direito (ITSVD) e do ventrículo esquerdo (ITSVE). Ocorreram dois óbitos transoperatórios no grupo de animais submetidos à oito minutos de parada circulatória total, devido a fibrilação ventricular. Frente aos óbitos ocorridos, conclui-se que o \"Inflow Occlusion\" foi exequível em cães sadios por sete minutos, sendo contra-indicado para períodos de oito minutos. Entretanto, estudos clínicos envolvendo a técnica em questão, precisam ser realizados para que a referida técnica possa ser aplicada, seguramente, em animais portadores de disfunções cardiovasculares. / Because of difficulties related to extracorporeal circulation in Veterinary Medicine, more interest occurred about circulatory arrest trough \"Inflow Occlusion\" technique to correct surgically some cardiac pathologies, such as semilunar valves stenosis. The aim of this study was to compare possible hemodynamic alterations that can occur in dogs submitted to circulatory arrest using \"Inflow Occlusion\" technique for different periods of time. Twelve mongrel dogs were used in this study divided in two groups, each one submitted to seven or eight minutes of total circulatory arrest. Hemodynamic evaluation was realized during four moments on transoperatory period. Oxygenation parameters were obtained trough pulmonary artery catheter data and hemogasometric analysis. Statistical difference between groups were found on M3 moment, five minutes after circulatory arrest, for mean arterial pressure (MAP), right ventricular systolic stroke index (RVSSI) and left ventricular systolic stroke index (LVSSI). There were two transoperatory deaths on group B, because ventricular fibrillation. Because these deaths, we can say that \"Inflow Occlusion\" can be realized up to seven minutes on healthy dogs and is contra-indicated for periods of eight minutes. However, clinical studies should be performed in order to use this technique routinely with safe in dogs presenting cardiovascular abnormalities.

Die vorliegende Arbeit setzt sich mit den funktionellen und strukturellen Veränderungen im Zentralnervensystem im Zusammenhang mit angeborenen Vitien und deren chirurgischer Behandlung mit Hilfe der extrakorporalen Zirkulation (EKZ) sowohl klinisch als auch tierexperimentell auseinander mit dem Ziel, neuroprotektive Strategien zu entwickeln. Wir haben mit den verfügbaren Methoden der Neuroüberwachung die charakteristischen Verläufe definiert und beschrieben. Zusätzlich wurden diese nicht-invasiven Methoden wie die Nahinfrarot-Spektroskopie sowohl klinisch als auch tierexperimentell validisiert. Es konnte jedoch gezeigt werden, dass diese Methoden eine zuverlässig signifikante globale Alteration in der Oxygenation und Perfusion anzeigen. Durch das Erarbeiten und die Charakterisierung des Verlaufs der Serumwerte des astroglialen Proteins S-100B wurde die klinische Wertigkeit genauer definiert. Es konnte klinisch und tierexperimentell gezeigt werden, dass die abnorm erhöhten Werte des S-100B im Serum von einem signifikanten diagnostischen Wert sind. Im Gegensatz dazu wurde die untergeordnete Rolle der Bestimmung von neuronalen Marker im Serum bestätigt. Durch die tierexperimentellen Arbeiten wurde gezeigt, dass die überwiegenden morphologischen Veränderungen nach EKZ im Gehirn in den Astrozyten und Gliazelen zu finden sind. Die neuronale Zelldegeneration war nach dem tiefhypothermen Kreislaufstillstand überwiegend in Form von hypoxischer Zellnekrose. Die apoptotische Zelldegeneration trat zellspezifisch im Gyrus Dentatus des Hippocampus auf. Vor allem konnte die bedeutende protektive Rolle der Hypothermie und der hypothermen Perfusion der EKZ demonstriert werden. Bei einer effektiven systemischen Kühlung an der EKZ könnte ein Kreislaufstillstand ohne signifikante neuronale Schädigungen überstanden werden. Die EKZ und der tiefhypotherme Kreislaufstillstand stellen an sich für das unreife Gehirn eine grobe nicht-physiologische Situation dar. Im Tiermodel könnte histologisch gezeigt werden, das die systemische Vorbehandlung mit Methylprednisolone keine protektive Wirkung hat. Obwohl eine signifikante Neuroprotektion durch Gabe von FK506 und Cyclosporin unter extremen Bedingungen der EKZ und tiefhypothermem Kreislaufstillstand erzielt wurde, bedarf es vor einer klinischen Anwendung weiterer tierexperimenteller und klinischer Überprüfungen. / The aim of our clinical and experimental studies was to evaluate functional and structural changes in the brain during corrective cardiac surgery using cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) in order to develop neuroprotective strategies. Using the available neurmonitoring methods such as the transcranial Doppler and near infrared spectroscopy (NIRS) characteristic changes in cerebral perfusion and oxygenation were defined and described according to the changes in hemodynamic parameters such perfusion pressure, temperature and flow rate. The diagnostic value of the astrocytic cell protein S100B was evaluated by measurement of the serum concentrations in infants and children with and without neurological complications. Additionally, the normal and abnormal release patterns were evaluated in experimental setting using an animal model of CPB and DHCA. According to the neuropathological assessment of the brain initial morphological changes were found predominantly in the astroglial cells. Systemic hypothermic perfusion on CPB before the induction of circulatory arrest period of 60 minutes was significantly protective. Ischemic neuronal injury in form of cell nekrosis was found in different brain region particularly after the prolongation of circulatory arrest time in deep hypothermia. The apoptotic cell death was found predominantly in the hippocampal region of the dentate gyrus. The routinely prophylactic systemic use steroid during cardiac surgery is not protective against ischemia and has been found to induce apoptosis in the hippocampus. In the same model the systemic pre-treatment with single high dose of Cyclosporin and FK506 decreased significantly the ischemic neuronal cell injury in different brain region. However, before clinical use further studies are necessary to optimise the dose and mode of application.

Hintergrund: Das akute Nierenversagen ist eine häufige Komplikation nach einer Herzoperation bei Neugeborenen, Säuglingen und Kleinkindern. Die Bedeutung der postoperativen Hämodynamik für eine Nierenschädigung ist gut bekannt, jedoch ist der Einfluß des kardiopulmonalen Bypasses und des tiefen hypothermen Kreislaufstillstandes weniger klar. Überdies gibt es Veränderungen der Blutviskosität während und nach der Herzoperation am kardiopulmonalen Bypass, welche die Nierenfunktion beeinflussen können. Aus diesem Grunde wurde der Einfluß der Blutviskosität am kardiopulmonalen Bypass und des tiefen hypothermen Kreislaufstillstandes auf die Nierenfunktion in dieser Patientengruppe untersucht. Methode: Untersucht wurden 44 Patienten mit einem Körpergewicht unter 10 kg, die am kardiopulmonalen Bypass operiert wurden. Von diesen erfolgte die Herzoperation bei 7 Patienten unter zusätzlicher Anwendung des tiefen hypothermen Kreislaufstillstandes. Bei allen Patienten wurden zu verschiedenen Zeitperioden Messungen zur Beschreibung der Nierenfunktion (Diurese, Kreatinin-Clearance und Gesamtprotein, Albumin, alpha-1-Mikroglobulin, Transferrin, IgG, N-Acetyl-beta-D-Glucosaminidase im Urin) und Bestimmungen der Blut- und Plasmaviskosität, der Erythrozytenaggregation und des kolloidosmotischen Druckes durchgeführt. Beide Gruppen wurden hinsichtlich des Einflusses des Kreislaufstillstandes auf die Nierenfunktion miteinander verglichen. Ergebnisse: Die während des kardiopulmonalen Bypasses im Zusammenhang mit einem erhöhten transglomerulären Filtrationsgradienten entstandene Polyurie und Proteinurie normalisierte sich innerhalb von 24 Stunden postoperativ. Die renale Ausscheidung von N-Acetyl-beta-D-Glucosaminidase und die erhöhte Natriumausscheidung zeigten zusätzlich eine tubuläre Schädigung an. Bei Hypothermie hatte die Plasmaviskosität einen deutlichen Einfluß auf die Blutviskosität, die während hypothermer Perfusion mit den im Urin gemessenen Werten von Albumin und N-Acetyl-beta-D-Glucosaminidase korrelierte. Die Patienten in der Kreislaufstillstandsgruppe hatten eine längere Bypasszeit und eine niedrigere minimale Körpertemperatur im Vergleich zu den Patienten ohne Kreislaufstillstand (p < 0,05). Diurese und Kreatinin-Clearance zeigten keine Differenzen zwischen beiden Gruppen. Während der Reperfusion wurde in der Kreislaufstillstandsgruppe signifikant mehr Albumin renal ausgeschieden als in der Vergleichsgruppe, ebenso Albumin und N-Acetyl-beta-D-Glucosaminidase nach dem kardiopulmonalen Bypass (p < 0,01). Schlußfolgerung: Die kardiopulmonale Bypassperfusion könnte eine Proteinurie und einen milden tubulären Schaden verursachen. Die Blutviskosität scheint dafür ein mitbestimmender Faktor zu sein und ist möglicherweise während hypothermer Perfusion wesentlich von der Plasmaviskosität abhängig. Es ist notwendig und wünschenswert anhand einer prospektiven Interventionsstudie den Einfluß der Blut- und Plasmaviskosität auf die postoperative Nierenfunktion zu untersuchen. Der tiefe hypotherme Kreislaufstillstand kann die Empfindlichkeit der Niere für einen Ischämie-Reperfusions-Schaden steigern. Obgleich die Befunde mild sind und keinen schweren ischämischen Nierenschaden anzeigen, sollte der durch den Kreislaufstillstand verursachte potentielle Nierenschaden für die Planung des chirurgischen Eingriffs bei Patienten mit angeborenen Herzfehlern als zusätzliches Risiko für ein akutes Nierenversagen mit in Betracht gezogen werden. / Background: Acute renal failure is a common complication after cardiopulmonary bypass in infants. Whereas it is well known that postoperative hemodynamics inflict acute renal failure, the influence of extracoporeal circulation on the kidney is less clear. Moreover, changes in blood viscosity occur during and after surgery, which may influence renal dysfunction. For this reason, the impact of blood viscosity during cardiopulmonary bypass and circulatory arrest on renal function was investigated. Methods: 44 patients weighting less than 10 kg operated on cardiopulmonary bypass were investigated, inclusive of 7 patients who additionally underwent circulatory arrest. In all patients analyses of renal function (diuresis, creatinine clearance, urinary total protein, albumin, alpha-1-microglobulin, transferrin, IgG, and N-acetyl-beta-D-glucosaminidase), blood, and plasma viscosity measurements, erythrocyte aggregation and colloid osmotic pressure were performed. Both groups were compared with regard to the impact of circulatory arrest on renal function. Results: Polyuria and proteinuria that appeared during cardiopulmonary bypass indicated an elevated transglomerular filtration gradient, which recovered within 24 hours. The appearance of N-acetyl-beta-D-glucosaminidase in the urine and elevated sodium excretion were additionally indicative of mild tubular damage. With hypothermia, plasma viscosity could had a major impact on the blood viscosity, which, during hypothermic perfusion, seemed to be related to proteinuria and N-acetyl-beta-D-glucosaminidase values. The patients of the circulatory arrest group had a longer bypass time and a lower body temperature in compare to the patients without circulatory arrest (p < 0.05). Diuresis and creatinine clearance revealed no differences between both groups. During reperfusion in the circulatory arrest group significantly more albumin were excreted as in the comparison group, likewise albumin and N-acetyl-beta-D-glucosaminidase after cardiopulmonary bypass (p < 0.01). Conclusions: Cardiopulmonary bypass perfusion could cause proteinuria and mild tubular damage. Blood viscosity may be one possible contributing factor, which in hypothermia may depend mainly on plasma viscosity. It is necessary and desirable to investigate the impact of blood, and plasma viscosity on postoperative renal function based on a prospective intervention study. The deep hypothermic circulatory arrest can increase the sensitivity of the kidney to an ischemia-reperfusion injury. Although the findings are mild and do not indicate severe ischemic renal damage, potential renal damage by deep hypothermic circulatory arrest should be taken into account for planning surgical procedures for congenital heart disease patients with additional risks of acute renal failure.