Transplantation of Reduced-Size Fatty Livers

缩小脂肪肝移植

基本信息

批准号：
8012891
负责人：
ZHI ZHONG
金额：
$ 3.18万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-22 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8012891
关键词：
3-nitrotyrosine 4 hydroxynonenal Accounting Acetylcysteine Adenoviruses Affect Antibodies Antioxidants Apoptosis Bilirubin Bromodeoxyuridine Calories Cell Cycle Cell Proliferation Cellular biology Clinic Complex Corn Oil Cryopreservation Cyclin D1 Cyclosporine Cysteine Cytosol Data Development Diet EGF gene Electron Spin Resonance Spectroscopy Energy Supply Enzymes Failure Fatty Liver Fatty acid glycerol esters Fluorescent Probes Free Radicals Freezing Fright Functional disorder Gene Delivery Genes Goals Graft Survival Growth Factor Harvest Hepatic Hepatic Mass Histology Hydrogen Peroxide Hypoxia Image Immunohistochemistry Immunosuppressive Agents Implant Incidence Infiltration Injury Interleukin-6 Lactated Ringer&apos s Solution Liver Liver Regeneration Living Donor Liver Transplantation Manganese Superoxide Dismutase Measurement Mediating Membrane Potentials Microcirculation Microscopy Mitochondria Mitochondrial Swelling Mitotic Modeling Molecular Biology Techniques NG-Nitroarginine Methyl Ester Natural regeneration Necrosis Nitrates Nitric Oxide Nitric Oxide Synthase Nitrites Nitrogen Obese Mice Outcome Oxidation-Reduction Oxygen Pathway interactions Permeability Peroxonitrite Pimonidazole Play Ploidies Production Proliferating Cell Nuclear Antigen Rattus Reactive Oxygen Species Reagent Respiratory Chain Role STAT3 gene Serum Severities Signal Transduction Pathway Solutions Source Spin Trapping Staining method Stains Stress Superoxide Dismutase Superoxides Techniques Testing Time Transplantation Triglycerides UCP2 protein University of Wisconsin-lactobionate solution Western Blotting Work adduct adverse outcome analog base choline deficient diet clinical application cytochrome c cytokine effective therapy feeding fluorexon fluorophore graft failure graft function human NOS2A protein immunocytochemistry implantation improved in vivo inhibitor/antagonist insight intravital video microscopy liver function liver transplantation mitochondrial dysfunction mitochondrial membrane non-alcoholic oil red O polyphenol prevent research study tetramethylrhodamine methyl ester transcription factor

项目摘要

DESCRIPTION (provided by applicant): Fatty livers are rarely used for partial liver transplantation due to the fear of primary non-function. These studies are to elucidate mechanisms and to develop strategies to prevent failure of fatty partial grafts. We will use reduced-size rat liver transplantation to test the unique hypotheses that steatosis and partial liver transplantation synergistically increase reactive oxygen and nitrogen species (ROS and RNS), inhibit mitochondrial function and suppress liver regeneration. In Aim 1, we will investigate if steatosis increases graft failure after partial liver transplantation. Fatty livers will be induced by high fat diet and choline-deficient diet, respectively, and reduced to 30% to 50% of original size. We will assess survival, liver function, and histology after transplantation as a function of steatosis, graft volume and cold storage time. In Aim 2, we will test the hypothesis that ROS and RNS are increased in high-fat diet-induced fatty partial grafts, leading to injury. ROS will be determined by the spin-trapping technique and immunohistochemistry for 4-hydroxynonenal and RNS will be assessed as nitrotyrosine and nitrite/nitrate levels. We expect that ROS and RNS production will be higher in fatty than lean partial grafts. We will also identify cellular sources of ROS and RNS using fluorescent intravital multiphoton microscopy. The effects of antioxidants (Ad-superoxide dismutase, N-acetylcysteine and polyphenols) and antinitrative therapies (nitric oxide synthase inhibitors) on survival and injury of fatty partial grafts will be evaluated. In Aim 3, effects of steatosis and graft volume on energy supply will be evaluated. Uncoupling protein 2, mitochondrial permeability transition and respiratory chain activity will be determined to elucidate the mechanisms of defective energy production, and effects of mitochondrial permeability transition inhibitors and antioxidant and antinitrative therapies on mitochondrial function will be evaluated. In Aim 4, the effects of steatosis and graft volume on liver regeneration will be assessed. Cell proliferation and factors regulating regeneration (cytokines, transcription factors, growth factors, and cell cycle related genes) will be evaluated. In addition, we will investigate if protection of mitochondria function and antioxidative and antinitrative therapies improve liver regeneration. Taken together, we expect that this work will provide fundamental new insights into the mechanisms underlying reduced-size fatty graft failure and develop mechanism-based strategies to increase the use and improve the outcome of fatty livers for partial liver transplantation in the clinic.

描述（由申请人提供）：由于担心原发性非功能，脂肪肝很少用于部分肝移植。这些研究旨在阐明机制并制定策略以防止脂肪部分移植物的失败。我们将使用减少大小的大鼠肝移植来测试脂肪变性和部分肝移植协同增加活性氧和氮种（ROS和RNS），抑制线粒体功能并抑制肝脏再生的独特假设。在AIM 1中，我们将调查脂肪变性是否会增加部分肝移植后移植物衰竭。高脂饮食和缺乏胆碱的饮食将诱导脂肪肝，并降低到原始大小的30％至50％。我们将评估移植后的生存，肝功能和组织学，这是脂肪变性，移植体积和冷藏时间的功能。在AIM 2中，我们将检验以下假设：高脂饮食诱导的脂肪部分移植物中ROS和RNS增加，导致损伤。 ROS将由4-羟基烯烯的自旋捕获技术和免疫组织化学确定，RN将评估为硝基酪氨酸和亚硝酸盐/硝酸盐水平。我们预计ROS和RNS的产量将比瘦的部分移植物高。我们还将使用荧光弹药多光子显微镜鉴定ROS和RN的细胞来源。评估抗氧化剂（酸氧化歧化酶，N-乙酰半胱氨酸和多酚）和抗硝化疗法（一氧化氮合酶抑制剂）对脂肪部分移植物的存活和损伤的影响。在AIM 3中，将评估脂肪变性和移植物量对能源供应的影响。将确定脱偶联蛋白2，线粒体通透性过渡和呼吸链活性，以阐明能量产生有缺陷的机制，以及线粒体通透性过渡抑制剂和抗氧化剂以及抗氧化疗法对线粒体功能的影响。在AIM 4中，将评估脂肪变性和移植物量对肝脏再生的影响。将评估细胞增殖和调节再生的因素（细胞因子，转录因子，生长因子和与细胞周期相关的基因）。此外，我们将研究线粒体功能以及抗氧化和抗毒性疗法是否可以改善肝脏再生。综上所述，我们预计这项工作将为减少大小的脂肪移植失败的机制提供基本的新见解，并制定基于机制的策略，以增加诊所中脂肪肝脏的脂肪肝脏的使用并改善脂肪肝脏的结果。