Ferrero Elisabetta, Scabini Silvia, Magni Elena, Foglieni Chiara, Belloni Daniela, Colombo Barbara, Curnis Flavio, Villa Antonello, Ferrero Maria Elena, Corti Angelo
Department Biological and Technological Research (DIBIT), San Raffaele H Scientific Institute, Via Olgettina 60, 20132 Milan, Italy.
FASEB J. 2004 Mar;18(3):554-6. doi: 10.1096/fj.03-0922fje. Epub 2004 Jan 20.
Elevated levels of circulating chromogranin A (CgA), a protein stored in the secretory granules of many neuroendocrine cells and neurons, have been detected in the blood of patients with neuroendocrine tumors or heart failure. The pathophysiological role of increased secretion of CgA is unknown. Using mice bearing subcutaneous tumors genetically engineered to secrete CgA in circulation, we have found that increased blood levels of this protein prevent vascular leakage induced by tumor necrosis factor-alpha (TNF) in the liver venous system. Structure-activity studies, carried out with CgA fragments administered to normal mice, showed that an active site is located within residues 7-57 of CgA. Accordingly, an anti-CgA antibody directed to residues 53-57 inhibited the effect of circulating CgA, either endogenously produced or exogenously administered, on liver vessels. Studies of the mechanism of action showed that CgA inhibits TNF-induced VE-cadherin down-regulation and barrier alteration of cultured endothelial cells, in an indirect manner. Other effectors, such as thrombin and vascular endothelial growth factor were partially inhibited by CgA N-terminal fragments in in vitro permeability assays. These findings suggest that circulating CgA could help regulate the endothelial barrier function and to protect vessels against TNF-induced plasma leakage in pathological conditions characterized by increased production of TNF and CgA, such as cancer or heart failure.
许多神经内分泌细胞和神经元的分泌颗粒中储存的一种蛋白质——循环嗜铬粒蛋白A(CgA)水平升高,已在神经内分泌肿瘤或心力衰竭患者的血液中被检测到。CgA分泌增加的病理生理作用尚不清楚。利用经基因工程改造可在循环中分泌CgA的皮下肿瘤小鼠,我们发现这种蛋白质血液水平的升高可预防肝静脉系统中由肿瘤坏死因子-α(TNF)诱导的血管渗漏。对正常小鼠给予CgA片段进行的结构-活性研究表明,一个活性位点位于CgA的7-57位残基内。因此,一种针对53-57位残基的抗CgA抗体抑制了循环CgA(无论是内源性产生还是外源性给予)对肝血管的作用。作用机制研究表明,CgA以间接方式抑制TNF诱导的培养内皮细胞中VE-钙黏蛋白下调和屏障改变。在体外通透性试验中,凝血酶和血管内皮生长因子等其他效应物被CgA N端片段部分抑制。这些发现表明,在以TNF和CgA产生增加为特征的病理状况(如癌症或心力衰竭)中,循环CgA可能有助于调节内皮屏障功能,并保护血管免受TNF诱导的血浆渗漏。