Glutathione and Lung Fibrosis

谷胱甘肽和肺纤维化

• 批准号: 7527229
• 负责人: RUI-MING LIU
• 金额: $ 34.9万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-25 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7527229
• 关键词: Acetylcysteine; Adenoviruses; Aerosols; Affect; Animal Model; Antioxidants; Attenuated; Bleomycin; Cells; Characteristics; Collagen; Cysteine; Deterioration; Disease; Fibroblasts; Fibrosis; Genetic; Glutamate-Cysteine Ligase; Glutathione; Glutathione Disulfide; Human; Immunoprecipitation; Inflammation; Inflammatory; Inflammatory Response; Intraperitoneal Injections; Knock-out; Knockout Mice; Label; Lung; Lung diseases; MAPK14 gene; MAPK8 gene; Matrix Metalloproteinases; Mediating; Modeling; Modification; Molecular; Mus; Oral Administration; Patients; Phosphoric Monoester Hydrolases; Phosphorylation; Physiologic pulse; Plasmin; Plasminogen; Plasminogen Activator; Plasminogen Activator Inhibitor 1; Play; Prevention strategy; Procollagen; Production; Proline; Protease Inhibitor; Protein Overexpression; Protein phosphatase; Proteins; Pulse taking; Reactive Oxygen Species; Respiratory physiology; Role; Saline; Signal Transduction; Small Interfering RNA; Staging; Structure of parenchyma of lung; Sulfhydryl Compounds; Supplementation; System; Techniques; Testing; Tetracycline; Tetracyclines; Therapeutic; Therapeutic Effect; Time; Tranexamic Acid; Transforming Growth Factor beta; Transforming Growth Factor-Beta Induced Protein IGH3; Transgenic Mice; Virus; Wild Type Mouse; autocrine; collagenase; computerized data processing; cytokine; day; design; fibrogenesis; inhibitor/antagonist; insight; mRNA Expression; novel; prevent; protein function; response

DESCRIPTION (provided by applicant): Lung fibrosis is a characteristic feature and terminal stage of many lung diseases with no efficacious treatment. The concentration of glutathione (GSH), the most abundant intracellular free thiol and an important antioxidant, is decreased in both experimental fibrosis and human fibrotic diseases. Importantly, aerosol or oral administration of GSH or N-acetylcysteine (NAC), a precursor of GSH, attenuates lung fibrosis in experimental fibrosis models and slows the deterioration of lung functions in patients with lung fibrotic diseases, indicating a potential therapeutic value of GSH/NAC for pulmonary fibrotic diseases. Nonetheless, the efficacy and mechanism underlying the therapeutic effects of GSH/NAC remains equivocal. Specifically, it is unclear whether GSH/NAC exerts its therapeutic effects by suppressing early-stage inflammatory response or whether it has direct antifibrotic activity and therefore can also block the progression of fibrosis in later stages of the disease. TGF-( is a most potent and ubiquitous profibrogenic cytokine. Our previous studies showed that TGF-( decreased GSH and increased reactive oxygen species (ROS) production in fibroblasts and in a lung fibrosis model while GSH/NAC supplementation inhibited TGF-(-stimulated collagen accumulation by blocking the expression of plasminogen activator inhibitor 1 (PAI-1), a protease inhibitor, and thus stimulating collagen degradation. Increasing GSH in lung ELF in tetracycline inducible lung specific glutamate cysteine ligase (GCL) transgenic mice also inhibits TGF-(-induced PAI-1 expression and lung fibrosis. Preliminary studies further show that TGF-( induces protein thiol modifications and inhibits the activity of JNK-directed phosphatases, associated with a stimulation of JNK/p38 phosphorylation while GSH blocks TGF-( activation of JNK and p38 MAPKs, which mediate TGF-( induction of PAI-1 expression. Therefore, we hypothesize that GSH/NAC can block fibrosis progression by attenuating TGF-('s fibrotic signaling and stimulating collagen degradation. Three specific aims are proposed that couple cell and animal models with pharmacologic and genetic approaches to elucidate the molecular mechanisms whereby GSH/NAC antagonizes TGF-(-induced lung fibrosis. Aim 1 will determine whether GSH/NAC can block the progression of late stage lung fibrosis induced by constitutively active TGF-( in GCL transgenic mice or NAC treated wild type mice. Aim 2 will determine whether GSH/NAC stimulates collagen degradation as well as the expression/activities of plasmin and the collagenases in TGF-( treated wild type and PAI-1 knockout mice. The bleomycin-induced lung fibrosis model will be used to further test our hypothesis. Aim 3 will focus on the molecular mechanisms whereby GSH/NAC antagonizes TGF-( fibrogenesis using mouse and primary human lung fibroblasts. Effect of GSH/NAC on TGF-(-induced thiol modifications and the activity of JNK/p38 directed phosphatases as well as the relationship between inhibition of the phosphatases and JNK/p38 activation/PAI-1 induction will be studied systematically. The results from these studies will provide new insight into the molecular mechanisms underlying TGF-('s fibrogenesis and GSH/NAC antifibrogenic effects, which will enable the design of more efficacious strategies for prevention and treatment of these fibroproliferative disorders. PROJECT NARRATIVE: Lung fibrosis is a characteristic feature and terminal stage of many lung diseases with no efficacious treatment. This project will test a novel hypothesis that glutathione, the most abundant intracellular free thiol and an important antioxidant, and N-acetyl cysteine (NAC), a precursor of GSH, can block fibrosis progression by attenuating the fibrogenic signals of TGF-(, a most potent and ubiquitous profibrogenic cytokine, and stimulating collagen degradation. The results from these studies will provide new insight into the molecular mechanisms underlying TGF-('s fibrogenesis and GSH/NAC antifibrogenic effects, which will enable the design of more efficacious strategies for prevention and treatment of these fibroproliferative disorders.

描述（由申请人提供）：肺纤维化是许多肺部疾病的特征和终末阶段，没有有效治疗。在实验性纤维化和人类纤维化疾病中，谷胱甘肽（GSH）的浓度是最丰富的细胞内游离硫醇和重要的抗氧化剂。重要的是，GSH或N-乙酰半胱氨酸（NAC）（GSH的前体）的气溶胶或口服给药可减弱实验性纤维化模型中的肺纤维化，并减慢肺纤维化疾病患者的肺功能恶化，表明GSH/NAC纤维纤维的潜在疗法值。尽管如此，GSH/NAC的治疗作用的效力和机制仍然是模棱两可的。具体而言，目前尚不清楚GSH/NAC是否通过抑制早期炎症反应或是否具有直接的抗纤维化活性来发挥其治疗作用，因此也可以阻止疾病后期纤维化的进展。 TGF-( is a most potent and ubiquitous profibrogenic cytokine. Our previous studies showed that TGF-( decreased GSH and increased reactive oxygen species (ROS) production in fibroblasts and in a lung fibrosis model while GSH/NAC supplementation inhibited TGF-(-stimulated collagen accumulation by blocking the expression of plasminogen activator inhibitor 1 (PAI-1), a protease抑制剂，从而刺激胶原蛋白降解。磷酸酶，与JNK/p38磷酸化的刺激相关，而GSH则阻止了TGF-（JNK和p38 MAPK的激活介导了TGF-（PAI-1表达的诱导诱导。 Three specific aims are proposed that couple cell and animal models with pharmacologic and genetic approaches to elucidate the molecular mechanisms whereby GSH/NAC antagonizes TGF-(-induced lung fibrosis. Aim 1 will determine whether GSH/NAC can block the progression of late stage lung fibrosis induced by constitutively active TGF-( in GCL transgenic mice or NAC treated wild type mice. Aim 2 will determine whether GSH/NAC刺激TGF-（处理过的野生型和PAI-1基因敲除小鼠。Bleyomycin诱导的肺纤维化模型的胶原蛋白降解以及纤溶酶和胶原酶的表达/活性。成纤维细胞。这些研究的结果将提供有关TGF基础的分子机制的新见解 - （纤维化和GSH/NAC/NAC抗纤维化效应，这将使您能够设计更有效的策略，以预防和治疗这些纤维增生性疾病。 glutathione, the most abundant intracellular free thiol and an important antioxidant, and N-acetyl cysteine (NAC), a precursor of GSH, can block fibrosis progression by attenuating the fibrogenic signals of TGF-(, a most potent and ubiquitous profibrogenic cytokine, and stimulating collagen degradation. The results from these studies will provide new insight into the TGF的分子机制 - （'s纤维发生和GSH/NAC抗纤维化作用，这将使设计更有效的策略，以预防和治疗这些纤维增生性疾病。