Pereira Thiago A, Syn Wing-Kin, Machado Mariana V, Vidigal Paula V, Resende Vivian, Voieta Izabela, Xie Guanhua, Otoni Alba, Souza Márcia M, Santos Elisângela T, Chan Isaac S, Trindade Guilherme V M, Choi Steve S, Witek Rafal P, Pereira Fausto E, Secor William E, Andrade Zilton A, Lambertucci José Roberto, Diehl Anna Mae
Division of Gastroenterology, Department of Medicine, Duke University Medical Center, Durham, NC 27710, U.S.A. Laboratório de Patologia Experimental, Centro de Pesquisas Gonçalo Moniz/FIOCRUZ, Salvador, BA 40296-710, Brazil.
Liver Regeneration and Repair Research Group, Institute of Hepatology, Foundation for Liver Research, London WC1E 6HX, U.K. Department of Surgery, Loyola University, Chicago, Maywood, IL 60153, U.S.A.
Clin Sci (Lond). 2015 Nov;129(10):875-83. doi: 10.1042/CS20150117. Epub 2015 Jul 21.
Schistosomiasis is a major cause of portal hypertension worldwide. It associates with portal fibrosis that develops during chronic infection. The mechanisms by which the pathogen evokes these host responses remain unclear. We evaluated the hypothesis that schistosome eggs release factors that directly stimulate liver cells to produce osteopontin (OPN), a pro-fibrogenic protein that stimulates hepatic stellate cells to become myofibroblasts. We also investigated the utility of OPN as a biomarker of fibrosis and/or severity of portal hypertension. Cultured cholangiocytes, Kupffer cells and hepatic stellate cells were treated with soluble egg antigen (SEA); OPN production was quantified by quantitative reverse transcriptase polymerase chain reaction (qRTPCR) and ELISA; cell proliferation was assessed by BrdU (5-bromo-2'-deoxyuridine). Mice were infected with Schistosoma mansoni for 6 or 16 weeks to cause early or advanced fibrosis. Liver OPN was evaluated by qRTPCR and immunohistochemistry (IHC) and correlated with liver fibrosis and serum OPN. Livers from patients with schistosomiasis mansoni (early fibrosis n=15; advanced fibrosis n=72) or healthy adults (n=22) were immunostained for OPN and fibrosis markers. Results were correlated with plasma OPN levels and splenic vein pressures. SEA-induced cholangiocyte proliferation and OPN secretion (P<0.001 compared with controls). Cholangiocytes were OPN (+) in Schistosoma-infected mice and humans. Liver and serum OPN levels correlated with fibrosis stage (mice: r=0.861; human r=0.672, P=0.0001) and myofibroblast accumulation (mice: r=0.800; human: r=0.761, P=0.0001). Numbers of OPN (+) bile ductules strongly correlated with splenic vein pressure (r=0.778; P=0.001). S. mansoni egg antigens stimulate cholangiocyte proliferation and OPN secretion. OPN levels in liver and blood correlate with fibrosis stage and portal hypertension severity.
血吸虫病是全球门静脉高压的主要病因。它与慢性感染期间发生的门静脉纤维化有关。病原体引发这些宿主反应的机制尚不清楚。我们评估了以下假设：血吸虫卵释放的因子直接刺激肝细胞产生骨桥蛋白（OPN），这是一种促纤维化蛋白，可刺激肝星状细胞转化为肌成纤维细胞。我们还研究了OPN作为纤维化和/或门静脉高压严重程度生物标志物的效用。用可溶性虫卵抗原（SEA）处理培养的胆管细胞、库普弗细胞和肝星状细胞；通过定量逆转录聚合酶链反应（qRTPCR）和酶联免疫吸附测定（ELISA）对OPN的产生进行定量；通过5-溴-2'-脱氧尿苷（BrdU）评估细胞增殖。用曼氏血吸虫感染小鼠6周或16周以诱导早期或晚期纤维化。通过qRTPCR和免疫组织化学（IHC）评估肝脏OPN，并将其与肝纤维化和血清OPN相关联。对曼氏血吸虫病患者（早期纤维化n = 15；晚期纤维化n = 72）或健康成年人（n = 22）的肝脏进行OPN和纤维化标志物免疫染色。结果与血浆OPN水平和脾静脉压力相关。SEA诱导胆管细胞增殖和OPN分泌（与对照组相比，P<0.001）。在感染血吸虫的小鼠和人类中，胆管细胞呈OPN阳性。肝脏和血清OPN水平与纤维化阶段相关（小鼠：r = 0.861；人类：r = 0.672，P = 0.0001）以及与肌成纤维细胞积累相关（小鼠：r = 0.800；人类：r = 0.761，P = 0.0001）。OPN阳性胆小管数量与脾静脉压力密切相关（r = 0.778；P = 0.001）。曼氏血吸虫卵抗原刺激胆管细胞增殖和OPN分泌。肝脏和血液中的OPN水平与纤维化阶段和门静脉高压严重程度相关。
