Liver Fibrosis, Inflammation & Oxidative Stress Markers

描述(由申请人提供)： 肝硬变是酗酒者死亡的主要原因。最近的实验性病理生理学研究提出了几种治疗方法，但其临床验证受到肝活检以确定疾病分期、治疗指征和结果记录的要求的阻碍。各种循环标志物已经被提出作为活组织检查的替代品。本研究的具体目的是验证这些标记物，利用超过2,000份连续的肝脏活检和相应的血液样本，在391年中对789名酗酒者进行长达6年的研究，包括每月测量碳水化合物缺乏的转铁蛋白，以证实自我报告的饮酒情况。这项最近进行的试验评估了多不饱和磷脂酰胆碱(PPC)，一种大豆提取物，对三分之一的患者合并丙型肝炎的酒精性肝病的效果。肝脏病理变化从脂肪肝到静脉周围和间隔纤维化以及肝硬变，并伴有不同程度的炎症。将确定这些损伤与下列拟议的循环纤维化标志物的相关性：层粘连蛋白、肌腱蛋白、前胶原型Tll和IV型和VI型胶原。还将测量降解纤维组织的酶的活性，如金属蛋白酶及其抑制物。此外，还将评估有可能预测肝脏疾病进展的肝脏参数，如细胞角蛋白。还将评估影响纤维化形成和炎症的细胞因子，如肿瘤坏死因子α和转化生长因子-β。一个主要的致病因素是由酒精诱导的细胞色素P450(CYP2E1)产生的自由基引起的氧化应激。因此，将评估氧化应激标志物(4-羟基壬烯醛、F2-异前列腺素和丙二醛)与肝脏中CYP2E1诱导程度的相关性。一旦得到验证，这些标志物可以在未来的研究中用于检测易受药物激活的药物的受试者。作为特定病变(如肝硬变)标记物的对照，我们将使用无特定肝病患者的相同标记物的值。总之，与详细的临床信息一起收集的连续肝活检和每月血样库提供了一个独特的机会，可以确定哪些循环标志物可以部分替代肝活检，从而优化酒精性肝损伤的诊断、预防和治疗。

DESCRIPTION (provided by applicant): Liver cirrhosis is a major cause of mortality in alcoholics. Recent experimental pathophysiotogic studies suggest several therapeutic approaches but their clinical verification is hampered by the requirement of liver biopsies to establish the stage of the disease, the therapeutic indications and documentation of outcome. Various circulating markers have been proposed as substitute for biopsies. The specific aim of the present study is to validate these markers, taking advantage of over 2,000 sequential liver biopsies and corresponding blood samples collected in 789 alcoholics studied for up to 6 years in VA Cooperative Study 391, including monthly measurements of carbohydrate deficient transferrin to substantiate the self-reported alcohol consumption. This recently conducted trial assessed the effects of polyunsaturated phosphatidylcholine (PPC), a soybean extract, on alcoholic liver disease, with superimposed hepatitis C in 1/3 of the patients. The liver pathology varied from fatty liver to perivenular and septal fibrosis and cirrhosis, with variable degrees of inflammation. Correlation of these lesions with the following proposed circulating markers of fibrosis will be determined: laminin, tenascin, procollagen type tll and collagen IV and VI. The activity of enzymes that degrade fibrotic tissue, such as metalloproteinases and their inhibitors, will also be measured. In addition, hepatic parameters with the potential to prognosticate progression of liver disorders, such as cytokeratins, will be assessed. Cytokines that affect fibrogenesis as well as inflammation, e.g. TNFalpha and TGF-beta will also be evaluated. One major pathogenic factor is oxidative stress resulting from free radicals generated by alcohol-induced cytochrome P450 (CYP2E1). Accordingly, the correlation of markers of oxidative stress (4-hydroxynonenal, F2-isoprostanes and malondialdehyde) with the degree of CYP2E1 induction in the liver will be evaluated. Once validated, these markers could serve in future studies to detect subjects vulnerable to drugs activated to toxins by CYP2E1. As controls for the markers of a specific lesion (such as cirrhosis), we will use the values of the same markers in patients without the specific liver disease. In conclusion, an available bank of sequential liver biopsies and monthly blood samples, collected in association with detailed clinical information, offers a unique opportunity to establish which circulating markers can serve as partial substitute for liver biopsies, thereby optimizing the diagnosis, prevention and treatment of alcoholic liver injury.