Moduration of the molecular chaperone activity to increase the safety of extended liver surgery in the damaged liver patients -- challenge by the induction of stress response and heat shock gene transfection --

1.非接触式隔离法评价最大肝切除量左三叶切除术中，只要缺血时间控制在90分钟以内，缺血时肝温保持在20℃左右，动物的存活期可持续5天以上。与异位手术相比，原位灌注中肝脏的冷却是不够的，因为肝脏被保持在体中而不切割肝门。但由于不需要重建肝动脉或胆管，因此该技术具有简化手术操作、缩短手术时间等优点。结果表明，该技术与常规肝切除术相比，可切除的肝体积无差异.分子伴侣活性调节的肝保护作用研究1）热休克预处理对细胞内环境的影响 ...更多信息 热休克预处理产生的热休克蛋白72参与了应激耐受的获得。我们发现热休克预处理可抑制NFκ-β的活化和TNF-α的产生。这些发现与热休克蛋白之间的关系还需要进一步研究。2）通过热休克基因转染肝脏获得应激耐受性我们建立了利用腺病毒将肌红蛋白和基质金属蛋白酶基因转染肝细胞的方法。3）诱导肝脏的应激反应除了诱导HSP 72的表达外，阿霉素还可抑制TNF-α的产生。反复热休克预处理可增加HSP 72蛋白表达，提高缺血耐受性。此外，我们发现，香叶基-香叶基-丙酮（GGA）的预处理加强应激反应，5分钟的热休克与GGA预处理产生缺血耐受的15分钟的热休克没有预处理相同的程度。少

1. Evaluation of maximally resectable volume of the liver by Non-touch isolation methodAnimal survival for more than 5 days was constantly obtained in the left trisegmentectomy with this technique as long as ischemic period was limited within 90 minutes and hepatic temperature during ischemia was kept at about 20℃. Cooling of the liver is not sufficient in in situ perfusion in comparison to the ex-situ operation because the liver is maintained in the corpus without cutting the hepatic hilum. However, this technique has big advantages in simplifying surgical technique and shortening operation time because neither the reconstruction of hepatic artery or bile duct is necessary. As a result, there was no difference in the resectable hepatic volume between this technique and conventional hepatic resection.2. Investigation on the liver protection by modurating molecular chaperon activity1)Analysis on the changes in intracellular milieu by heat shock preconditioningWe have already reported th … More at HSP72 was produced by heat shock preconditioning and this protein was involved in the acquisition of stress tolerance. Here, we found that the activation of NFκ-β and the production of TNF-α were suppressed by heat shock preconditioning. Further investigation is needed to know the relationship between these findings and heat shock proteins.2)Acquisition of stress tolerance by heat shock gene transfectlon to the liverWe established the method to transfect the myoglobin and matrix metalloprotease gene into hepatocytes using adenovirus. We are now studying how to transfect HSP 72 in hepatocytes by this method.3)Induction of stress response of the liverIn addition to the HSP72 induction, inhibition of TNF-α production was involved in the stress response to the treatment by Doxorubicin. We also found that repeated heat shock preconditioning increased the expression of HSP72 protein and ischemic tolerance. Moreover, we revealed that the pretreatment of geranyl-geranyl-acetone (GGA) intensified stress response and that 5-minute heat shock with GGA pretreatment produced ischemic tolerance to the same extent as 15-minute heat shock without pretreatment. Less