Infusion of human amniotic epithelial and mesenchymal stromal stem cells as a new therapeutic option for portal hypertension and cirrhosis

Background and aims. Portal hypertension is the main consequence of cirrhosis, responsible for the complications defining clinical decompensation. The only cure for decompensated cirrhosis is liver transplantation, but it is a limited resource and opens the possibility of regenerative therapy. We investigated the potential of human amniotic membrane-derived mesenchymal stromal (hAMSCs) and epithelial (hAECs) stem cells for the treatment of portal hypertension and chronic liver disease. Methods. In vivo: hAMSCs and hAECs were isolated from human amniotic membranes. Cirrhotic rats with ascites (chronic CCl4 inhalation) received 4x10e6 hAMSCs, 4x10e6 hAECs, or vehicle (NaCl 0.9%) (intraperitoneal; n=10 per group). After 2-week we analyzed: a) portal pressure (PP) and liver microcirculatory function; b) liver sinusoidal endothelial (LSECs) and hepatic stellate (HSCs) cells phenotype; c) hepatic fibrosis, inflammation and hepatic function. In vitro: HSCs isolated from CCl4-cirrhotic rats were co-cultured with hAMSCs, hAECs or vehicle for 24h. RNA profile was analyzed by RNAseq. Results. Cirrhotic rats receiving hAMSCs or hAECs had significantly lower PP than vehicle-treated animals, together with improved liver microcirculatory function. This hemodynamic amelioration was associated with improvement in LSECs capillarization and HSCs de-activation, although hepatic collagen/fibrosis was not significantly reduced. Rats that received placenta derived stem cells had markedly reduced hepatic inflammation and oxidative stress. Finally, liver function tests significantly improved in rats receiving hAMSCs. In vitro experiments confirmed HSCs de-activation when co-cultured with stem cells. Conclusion. This pre-clinical study shows that infusion of human amniotic stem cells effectively decreases PP by ameliorating liver microcirculation, suggesting that it may represent a new treatment option for advanced cirrhosis with portal hypertension.研究背景与目的。门静脉高压（portal hypertension）是肝硬化（cirrhosis）的主要并发症，也是引发临床失代偿（clinical decompensation）的核心因素。失代偿期肝硬化的唯一根治手段为肝移植（liver transplantation），但该资源有限，因此再生治疗（regenerative therapy）成为潜在的替代方案。本研究探讨了人羊膜来源间充质基质干细胞（human amniotic membrane-derived mesenchymal stromal cells, hAMSCs）与上皮干细胞（human amniotic membrane-derived epithelial cells, hAECs）用于治疗门静脉高压与慢性肝病的潜力。 研究方法。体内实验：从人羊膜中分离hAMSCs与hAECs。采用慢性四氯化碳（CCl4）吸入法构建伴腹水的肝硬化大鼠模型，分别向模型大鼠腹腔注射4×10^6个hAMSCs、4×10^6个hAECs或0.9%氯化钠（NaCl 0.9%）溶剂，每组10只。造模2周后开展以下检测：a) 门静脉压力（portal pressure, PP）与肝脏微循环功能；b) 肝窦内皮细胞（liver sinusoidal endothelial cells, LSECs）与肝星状细胞（hepatic stellate cells, HSCs）表型；c) 肝纤维化、炎症反应与肝功能。 体外实验：从CCl4诱导的肝硬化大鼠体内分离HSCs，将其分别与hAMSCs、hAECs或溶剂共培养24小时，通过RNA测序（RNAseq）分析转录组表达谱。 研究结果。接受hAMSCs或hAECs治疗的肝硬化大鼠，其门静脉压力显著低于溶剂对照组，同时肝脏微循环功能得到改善。该血流动力学改善与LSECs毛细血管化程度减轻及HSCs活化受抑相关，但肝胶原/纤维化水平未出现显著降低。接受胎盘来源干细胞治疗的大鼠，其肝脏炎症与氧化应激水平显著缓解。此外，接受hAMSCs治疗的大鼠肝功能指标得到显著改善。体外实验证实，与干细胞共培养可有效抑制HSCs活化。 研究结论。这项临床前研究表明，输注人羊膜来源干细胞可通过改善肝脏微循环有效降低门静脉压力，提示其有望成为合并门静脉高压的晚期肝硬化的新型治疗选择。