Mechanism of oxidative/nitrosative stress and inflammation-induced tissue injury

氧化/亚硝化应激与炎症引起的组织损伤的机制

• 批准号: 10266450
• 负责人: PAL PACHER
• 金额: $ 222.02万
• 依托单位: NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10266450
• 关键词: Acquired Immunodeficiency Syndrome; Acute; Affect; Aging; Agonist; Alcoholic Liver Diseases; Alcoholic steatohepatitis; Alcohols; Animal Disease Models; Animal Diseases; Animals; Antibody Therapy; Atherosclerosis; Attenuated; Bacterial Infections; Biological Availability; CNR2 gene; Cardiac; Cardiac development; Cardiomyopathies; Cardiovascular Diseases; Cardiovascular Physiology; Cardiovascular system; Cell Death; Cells; Characteristics; Chronic; Cirrhosis; Clinical; Clinical Research; Collaborations; Complex; Complication; Coupled; Detection; Development; Disease; Disease model; E-Selectin; Echocardiography; Encephalopathies; Endocannabinoids; Enzymes; Fibrosis; Functional disorder; Future; Heart; Heart failure; Hepatic; Hepatopulmonary Syndrome; Hepatorenal Syndrome; Hypotension; IL6 gene; Impairment; Inferior Vena Cava Occlusion; Infiltration; Inflammation; Inflammatory; Injury; Injury to Kidney; Intercellular adhesion molecule 1; Interleukin-1; Kidney; Kidney Diseases; Kidney Failure; Laboratories; Leukocytes; Ligation; Lipids; Liver; Liver Failure; Liver Fibrosis; Liver Regeneration; Liver diseases; Metabolic; Microcirculation; Modeling; Molecular; Monoclonal Antibodies; Mus; Myocardial; Myocardial Ischemia; Myocardial dysfunction; NAPE-PLD; Names; Nitrates; Oxidative Stress; Pathology; Pathway interactions; Patients; Performance; Peripheral; Poly(ADP-ribose) Polymerases; Portal Hypertension; Rattus; Receptor Activation; Reperfusion Injury; Reporting; Resistance; Role; Serum; Signal Transduction; Stress; Syndrome; TNF gene; Testing; Therapeutic; Tissues; Tubular formation; Vascular Cell Adhesion Molecule-1; Vascular Diseases; Vasoconstrictor Agents; Vasodilation; activity-based protein profiling; alcohol use disorder; attenuation; bile duct; body system; cardiovascular injury; chemokine; chronic liver disease; chronic liver injury; clinically relevant; cytokine; emotional behavior; endocannabinoid signaling; endogenous cannabinoid system; heart function; humanized mouse; improved; indexing; inhibitor/antagonist; interest; kidney dysfunction; liver development; liver inflammation; liver injury; mortality; mouse model; neuroinflammation; new therapeutic target; nitrosative stress; novel; novel therapeutic intervention; outcome forecast; pre-clinical; receptor expression; regenerative; tissue injury; tool; vascular inflammation

Interplay of oxidative/nitrative stress, inflammation with the endocannabinoid system (ES) in tissue injury and inflammation. Recently it has become clear that there are very important interactions between various organ systems, which may promote cardiovascular disease and/or development of various severe cardiovascular complications in certain diseases (e.g. chronic liver and kidney diseases). We have been particularly interested in the interactions of liver with the cardiovascular system and kidneys during chronic hepatic inflammation and injury. It is well known that chronic liver diseases are among the top contributors to mortality worldwide and that patients with severe liver disease (both acute and chronic) often develop different cardiovascular complications including vasculopathies, portal hypertension, peripheral splanchnic vasodilation resistant to vasoconstrictors, hepatopulmonary syndrome, encephalopathy, hepatorenal syndrome and cirrhotic cardiomyopathy, just naming a few, which ultimately largely contribute to the increased mortality observed in this patient group. In a recent study we aimed to investigate the potential interplay of cardiovascular system and liver during chronic liver disease using a well-established model of liver inflammation and fibrosis induced by bile duct ligation (BDL) in mice. To determine cardiovascular function, we used multiple approaches including echocardiography and intracardiac P-V analysis, among others. Consistently with previous studies, we confirmed the development of liver inflammation, impaired microcirculation and fibrosis 2 weeks following BDL in mice. In parallel we found diastolic and systolic dysfunction in mice and hypotension. The systolic dysfunction became evident only when the load-independent indices of LV performance were evaluated by P-V approach following manipulation of the cardiac load by transient vena cava inferior occlusions. To explore the impact of attenuation of liver inflammation on the development of cardiac dysfunction and myocardial pathology, we used the selective CB2R agonist HU910. Consistently with previous reports, CB2R activation in the liver effectively reduced BDL-induced liver fibrosis, leukocyte infiltration, and the associated hepatic oxidative. Importantly, HU910 treatment also restored the hepatic microcirculatory dysfunction in BDL animals. The above changes were supported by significantly lower levels of ALT and ALP and markedly reduced expression levels of multiple proinflammatory cytokines/chemokines (IL1, IL6, MIP1, TNF) and markers of vascular inflammation (E-selectin, VCAM-1, ICAM-1), and by significantly lower serum TNF levels in HU910-treated BDL mice. In addition to improving liver inflammation/fibrosis and microcirculation, the treatment also attenuated myocardial cellular and vascular inflammation, oxidative/nitrative stress, and markers of cardiomyopathy resulting in improved cardiac function in hearts of BDL mice. In summary, we demonstrate that BDL-induced advanced liver fibrosis is a suitable mouse model to study the pathophysiology of hepatic/cirrhotic cardiomyopathy at a preclinical level as it resembles the characteristics of the clinical syndrome seen in patients. Our results indicate that hepatic cardiomyopathy is characterized by myocardial systolic and diastolic dysfunction coupled with significant changes in macrovascular and microvascular function. Our results also show that the liverheart inflammatory axis has a pivotal pathophysiological role in the development of hepatic cardiomyopathy. In addition to affecting cardiovascular function, liver disease may also profoundly interact with other organ systems. Hepatorenal syndrome (HRS) is a very frequent and potentially lethal complication of acute and chronic liver failure (e.g. induced by alcoholic steatohepatitis, cirrhosis, etc.) and is an important predictor of short-term mortality with poor prognosis. Previously we established a clinically relevant mouse model of hepatorenal syndrome and provided evidence on a potential mechanism of the microcirculatory collapse resulting in kidney failure in HRS. More recently, using this model we explored the therapeutic potential of targeting the non-psychoactive cannabinoid-2 receptor (CB2R). We found that liver injury induced by BDL markedly increased markers of kidney injury, triggered pronounced histopathological alterations in the kidney with tubular injury and early fibrotic changes, markedly enhanced renal inflammation and oxidative/nitrative stress and vascular inflammation resulting in dramatically reduced microcirculatory collapse and kidney dysfunction. This was accompanied by increased CB2R expression in the kidney and liver tissues of diseased animals. A selective CB2R agonist, HU-910, markedly decreased numerous markers of kidney injury, oxidative stress inflammation. HU-910 also attenuated vascular inflammation and improved the impaired renal microcirculation in BDL-operated mice, likewise the BDL-induced liver pathology. Our future studies will focus on the development of improved tools for cannabinoid 2 receptor (CB2R) detection in tissues and cell and on development of novel more selective CB2R agonists with improved selectivity and bioavailability. Our collaborative studies with Dr. Yuri Persidsky have also been exploring the role of CB2R in AIDS using humanized mouse models of the disease. In recent collaborative studies with Dr. Van der Stelt we described a NAPE-PLD inhibitor that modulates emotional behavior in mice and also tested the feasibility of the use of activity-based protein profiling and lipidomic in ischemic hearts. Our impending studies will also focus on the understanding of the mechanisms of the activation of the endocannabinoid system during tissue injury and on the further elucidation of the role of endocannabinoid system (particularly focusing on the endocannabinoid metabolizing enzymes and CB2R in collaboration with Drs. Cravatt, Van der Stelt, Mechoulam and Kunos) in various models of liver disease, cardiomyopathy and nephropathy. Role of oxidative/nitrosative stress, inflammation, metabolic dysregulation in tissue injury Recently, in collaboration with Drs. Lohoff and Koob we discovered that inhibition of PCSK9 in rats with a therapeutically approved monoclonal antibody for the treatment of atherosclerosis may represent a novel therapeutic approach for alcoholic liver disease by attenuating hepatic inflammation and steatosis. In a small-scale clinical study, we plan to explore the translatability of this approach and plan to include numerous cardiovascular endpoints. In a collaborative study with Dr. Gao describing a new mouse model we demonstrated that acute-on-chronic liver injury or bacterial infection is associated with impaired liver regeneration due to a shift from a pro-regenerative to an anti-regenerative pathway. IL-22Fc therapy attenuated this shift and attenuated bacterial infection, suggesting that IL-22Fc may have therapeutic potential for ACLF treatment. Our future collaborative studies with Drs. Kunos, Koob, Gao, and Falk will also explore the role of oxidative/nitrosative stress, inflammation, lipid endocannabinoid signaling and cell death in various new rat and mouse models of alcohol use disorders.

在组织损伤和炎症中的氧化/硝化应激，炎症与内源性大麻素系统（ES）的相互作用。 最近，很明显，各种器官系统之间存在非常重要的相互作用，这可能会促进心血管疾病和/或某些疾病中各种严重心血管并发症的发展（例如慢性肝脏和肾脏疾病）。在慢性肝发炎和损伤期间，我们对肝脏与心血管系统和肾脏的相互作用特别感兴趣。 It is well known that chronic liver diseases are among the top contributors to mortality worldwide and that patients with severe liver disease (both acute and chronic) often develop different cardiovascular complications including vasculopathies, portal hypertension, peripheral splanchnic vasodilation resistant to vasoconstrictors, hepatopulmonary syndrome, encephalopathy, hepatorenal syndrome and肝硬化心肌病，只是命名了一些，最终在很大程度上导致了该患者组中观察到的死亡率增加。 在最近的一项研究中，我们旨在利用胆汁管结扎（BDL）在小鼠中诱导的肝脏炎症和纤维化模型来研究慢性肝病期间心血管系统和肝脏的潜在相互作用。为了确定心血管功能，我们使用了多种方法，包括超声心动图和心内P-V分析等。 与以前的研究一致，我们证实了小鼠BDL后2周的肝脏炎症，微循环受损和纤维化的发展。同时，我们发现小鼠和低血压中的舒张期和收缩功能障碍。仅当通过瞬态静脉腔内闭塞操纵心脏负荷后，通过P-V方法评估LV性能的载荷性能指数，仅在lv性能的载荷性能指数时才变得明显。 为了探索肝脏炎症衰减对心脏功能障碍和心肌病理发展的影响，我们使用了选择性的CB2R激动剂HU910。与先前的报道一致，肝脏中的CB2R激活有效地降低了BDL诱导的肝纤维化，白细胞浸润和相关的肝氧化性。重要的是，HU910治疗还恢复了BDL动物中肝微循环功能障碍。上述变化由ALT和ALP的水平显着降低，并显着降低了多种促炎细胞因子/趋化因子（IL1，IL6，MIP1，TNF）的表达水平，以及血管炎症的标记（E-塞染料，VCAM-1，ICAM，ICAM，ICAM，ICAM-1）的表达水平，以及高血清TNF TNF TNF水平Intnf水平Inhu910-bd bd tnf水平。除了改善肝脏炎症/纤维化和微循环外，该治疗还减弱了心肌和血管炎症，氧化/硝化应激以及心肌病的标志物，从而改善了BDL小鼠心脏中心脏功能。 总而言之，我们证明了BDL诱导的晚期肝纤维化是一种合适的小鼠模型，可在临床前研究肝/肝硬化心肌病的病理生理学，因为它类似于患者中临床综合症的特征。我们的结果表明，肝心肌病的特征是心肌收缩和舒张功能障碍以及大血管和微血管功能的显着变化。我们的结果还表明，肝脏炎症轴在肝心肌病的发展中具有关键的病理生理作用。 除了影响心血管功能外，肝病还可能与其他器官系统深刻相互作用。肝烯综合征（HRS）是一种非常频繁的急性和慢性肝衰竭并发症（例如，通过酒精性脂肪性肝炎，肝硬化等诱导），是预后不良的短期死亡率的重要预测指标。以前，我们建立了与肝癌综合征的临床相关小鼠模型，并提供了微循环塌陷的潜在机制的证据，导致HRS肾衰竭。最近，使用该模型，我们探讨了靶向非精神活性大麻素-2受体（CB2R）的治疗潜力。我们发现，BDL引起的肝损伤显着增加了肾脏损伤的标记，引发了肾脏损伤和早期纤维化变化的明显组织病理学改变，显着增强了肾脏炎症以及氧化/硝化应激以及在显着减轻的微循环次数胶水胶水胶水和肾脏dysdydysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdysdys dysfuncount中。这伴随着患病动物的肾脏和肝组织中CB2R表达的增加。选择性CB2R激动剂HU-910显着降低了肾脏损伤，氧化应激炎症的许多标记。 HU-910还减轻了血管炎症，并改善了BDL锻炼小鼠的肾脏微循环受损，同样是BDL诱导的肝脏病理学。 我们未来的研究将集中于在组织和细胞中开发改进的大麻素2受体（CB2R）检测工具，以及具有提高选择性和生物利用度的新型更具选择性的CB2R激动剂的开发。我们与Yuri Persidsky博士的合作研究也一直在探索CB2R在使用该疾病的人源化小鼠模型中的艾滋病中的作用。 在最近与Van der Stelt博士的合作研究中，我们描述了一种NAPE-PLD抑制剂，该抑制剂调节小鼠的情绪行为，并测试了在缺血性心脏中使用基于活性的蛋白质分析和脂质组学的可行性。我们迫在眉睫的研究还将集中在组织损伤过程中内源性大麻素系统激活机制的理解以及进一步阐明内源性大麻素系统的作用（尤其是专注于内源性大麻素的代谢酶和CB2R，与Drs的Cravatt，van der stelt，Mechoul stelt，Mechoul and knode，Mechoul and Kuny and knode and knode and knode，Mechoul and knode，Mechoul and knode and knode and knode。肾病。 氧化/亚硝化应激，炎症，代谢失调在组织损伤中的作用 最近，与Drs合作。 Lohoff和Koob我们发现，对治疗性动脉粥样硬化治疗的治疗批准的单克隆抗体的PCSK9抑制PCSK9可能会通过减弱肝炎症和脂肪症来治疗酒精性肝病的新型治疗方法。在一项小型临床研究中，我们计划探索这种方法的转换性，并计划包括许多心血管终点。 在与GAO博士的一项合作研究中，我们描述了一种新的小鼠模型，我们证明，由于从促增生向抗新途径的转变，急性智力肝损伤或细菌感染与肝脏再生受损有关。 IL-22FC治疗减弱了这种转移并减弱细菌感染，这表明IL-22FC可能具有ACLF治疗的治疗潜力。 我们与Drs的未来合作研究。 Kunos，Koob，Gao和Falk还将探索各种新的大鼠酒精使用障碍大鼠模型中氧化/亚硝化应激，炎症，脂质内源性内源性信号传导和细胞死亡的作用。