Exosomes from Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Stromal Cells (hiPSC-MSCs) Protect Liver against Hepatic Ischemia/Reperfusion Injury via Activating Sphingosine Kinase and Sphingosine-1-Phosphate Signaling Pathway
Exosomes from Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Stromal Cells (hiPSC-MSCs) Protect Liver against Hepatic Ischemia/Reperfusion Injury via Activating Sphingosine Kinase and Sphingosine-1-Phosphate Signaling Pathway
复制标题
来自人诱导多能干细胞衍生的间充质基质细胞 (hiPSC-MSC) 的外泌体通过激活鞘氨醇激酶和 1-磷酸鞘氨醇信号通路保护肝脏免受肝缺血/再灌注损伤
DOI：
10.1159/000480533
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Chen, Xiaosong
中科院分区：
文献类型：
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作者：
Du, Yingdong;Li, Dawei;Chen, Xiaosong
背景/目标：本研究旨在探讨人诱导多能干细胞来源的间充质基质细胞（hiPSC-MSCs-Exo）产生的exosomes对肝脏缺血再灌注（I/R）损伤的影响及其机制。方法：从hiPSC-MSCs中分离出外泌体，并进行生化和生物药理学表征。将hiPSC-MSCs-Exo经下腔静脉注入小鼠脑缺血再灌注损伤模型，观察其治疗效果。检测血清转氨酶（谷草转氨酶（AST）和丙氨酸转氨酶（ALT））水平及组织学变化。使用原代肝细胞和人肝细胞系HL 7702来测试外泌体是否可以在体外诱导肝细胞增殖。另外，通过免疫组织化学和Western blot检测增殖标志物（增殖细胞核抗原，PCNA;磷酸化组蛋白-H3，PHH 3）的表达水平。采用SK抑制剂（SKI-II）和S1 P1受体拮抗剂（VPC 23019）研究鞘氨醇激酶和鞘氨醇-1-磷酸依赖性通路在hiPSC-MSCs-Exo对肝细胞的影响中的作用。结果：hiPSCs经高效诱导分化为具有典型MSC特征的hiPSC-MSCs。hiPSC-MSC-Exo的直径范围为100至200 nm，并表达外泌体标志物（阿利克斯、CD 63和CD 81）。hiPSC-MSCs-Exo给药后，缺血/再灌注损伤模型中肝细胞坏死和肝窦充血明显受到抑制，组织病理学评分较低。与对照组相比，治疗组中肝细胞损伤标志物AST和ALT的水平显著降低，并且在hiPSC-MSC-Exo施用后，增殖标志物（PCNA和PHH 3）的表达水平被极大地诱导。hiPSC-MSCs-Exo还能诱导原代肝细胞和HL 7702细胞增殖，并呈剂量依赖性。结果发现hiPSC-MSCs-Exo可直接与靶肝细胞或HL 7702细胞融合，并能增加鞘氨醇激酶活性和1-磷酸鞘氨醇（S1 P）合成。此外，SK 1或S1 P1受体的抑制完全消除了hiPSC-MSC-Exo对肝细胞的保护和增殖作用，无论是在体外还是在体内。结论：结果表明hiPSC-MSCs-Exo可通过激活肝细胞鞘氨醇激酶和鞘氨醇-1-磷酸途径减轻肝I/R损伤，促进细胞增殖。这些发现代表了一种新的机制，可能有助于肝再生，并对急性肝病的新治疗方法具有重要意义。(C)2017作者由S.发布Karger AG，巴塞尔
Background/Aims: This study aimed to evaluate the effects of exosomes produced by human-induced pluripotent stem cell-derived mesenchymal stromal cells (hiPSC-MSCs-Exo) on hepatic ischemia-reperfusion (I/R) injury, as well as the underlying mechanisms. Methods: Exosomes derived from hiPSC-MSCs were isolated and characterized both biochemically and biophysically. hiPSC-MSCs-Exo were injected systemically into a murine ischemia/reperfusion injury model via the inferior vena cava, and then the therapeutic effects were evaluated. The serum levels of transaminases (aspartate aminotransferase (AST) and alanine aminotransferase (ALT), as well as histological changes were examined. Primary hepatocytes and human hepatocyte cell line HL7702 were used to test whether exosomes could induce hepatocytes proliferation in vitro. In addition, the expression levels of proliferation markers (proliferation cell nuclear antigen, PCNA; Phosphohistone-H3, PHH3) were measured by immunohistochemistry and Western blot. Moreover, SK inhibitor (SKI-II) and S1P1 receptor antagonist (VPC23019) were used to investigate the role of sphingosine kinase and sphingosine-1-phosphate-dependent pathway in the effects of hiPSC-MSCs-Exo on hepatocytes. Results: hiPSCs were efficiently induced into hiPSC-MSCs that had typical MSC characteristics. hiPSC-MSCs-Exo had diameters ranging from 100 to 200 nm and expressed exosome markers (Alix, CD63 and CD81). After hiPSC-MSCs-Exo administration, hepatocyte necrosis and sinusoidal congestion were markedly suppressed in the ischemia/reperfusion injury model, with lower histopathological scores. The levels of hepatocyte injury markers AST and ALT were significantly lower in the treatment group compared to control, and the expression levels of proliferation markers (PCNA and PHH3) were greatly induced after hiPSC-MSCs-Exo administration. Moreover, hiPSC-MSCs-Exo also induced primary hepatocytes and HL7702 cells proliferation in vitro in a dose-dependent manner. We found that hiPSC-MSCs-Exo could directly fuse with target hepatocytes or HL7702 cells and increase the activity of sphingosine kinase and synthesis of sphingosine-1-phosphate (S1P). Furthermore, the inhibition of SK1 or S1P1 receptor completely abolished the protective and proliferative effects of hiPSC-MSCs-Exo on hepatocytes, both in vitro and in vivo. Conclusions: Our results demonstrated that hiPSC-MSCs-Exo could alleviate hepatic I/R injury via activating sphingosine kinase and sphingosine-1-phosphate pathway in hepatocytes and promote cell proliferation. These findings represent a novel mechanism that potentially contributes to liver regeneration and have important implications for new therapeutic approaches to acute liver disease. (C) 2017 The Author(s) Published by S. Karger AG, Basel