Hematopoietic stem cells and liver regeneration: Differentially acting hematopoietic stem cell mobilization agents reverse induced chronic liver injury
Hematopoietic stem cells and liver regeneration: Differentially acting hematopoietic stem cell mobilization agents reverse induced chronic liver injury
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DOI：
10.1016/j.bcmd.2014.05.003
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发表时间：
2014-09-01
影响因子：
2.3
通讯作者：
Yannaki, Evangelia
中科院分区：
文献类型：
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作者：
Tsolaki, Eleftheria;Athanasiou, Evangelia;Yannaki, Evangelia
在肝损伤的情况下，骨髓（BM）可以作为促进肝再生的细胞来源。目前可用的造血干细胞（HSC）动员剂，比较测试肝纤维化的愈合潜力。用粒细胞集落刺激因子（G-CSF）、普乐沙福或G-CSF +普乐沙福动员先前用绿色荧光蛋白BM重建的四氯化碳（CCl 4）损伤小鼠。肝纤维化、星状细胞活化和卵圆干细胞频率分别用Gomori法和α-平滑肌肌动蛋白和细胞角蛋白-19免疫组化法测定。用ELISA和免疫组化法检测血管生成。采用实时定量PCR测定肝脏过氧化物酶体激活受体γ（PPAR-gamma）、白细胞介素-6（IL-6）和肿瘤坏死因子α（TNF α）的mRNA水平。通过双重免疫荧光追踪BM衍生的细胞。在transwell培养系统中测定动员的HSC向损伤肝脏的自发迁移及其细胞因子分泌谱。无论是单一的代理动员或代理的组合显着改善肝损伤，减少纤维化和恢复异常的血管网络动员小鼠的肝脏相比，四氯化碳-只有小鼠。尽管G-CSF +普乐沙福产生的循环HSC数量显著高于其他药物，但所有动员小鼠的纤维化减少程度相似。各种动员的HSC的肝脏归巢潜力在药剂之间不同。延长的G-CSF治疗提供了最高的抗纤维化作用超过所有测试的方式，诱导肝干细胞的增殖和减少肝脏炎症。尽管普乐沙福动员的HSC肝脏归巢能力降低，但主要通过增加肝脏PPAR-gamma和VEGF表达逆转纤维化。在所有组中，BM衍生的成熟肝细胞以及肝脏定向BM干细胞仅在低频率下检测到，进一步支持替代机制而不是直接HSC作用调节肝脏恢复的概念。总体而言，我们的数据表明，G-CSF、Plerixafor和G-CSF + Plerixafor在伤口愈合过程中的作用不同，最终提供了有效的抗纤维化作用。(C)2014爱思唯尔公司All rights reserved.
Bone marrow (BM) could serve as a source of cells facilitating liver repopulation in case of hepatic damage. Currently available hematopoietic stem cell (HSC) mobilizing agents, were comparatively tested for healing potential in liver fibrosis. Carbon tetrachloride (CCl4)-injured mice previously reconstituted with Green Fluorescent Protein BM were mobilized with Granulocyte-Colony Stimulating Factor (G-CSF), Plerixafor or G-CSF + Plerixafor. Hepatic fibrosis, stellate cell activation and oval stem cell frequency were measured by Gomori and by immunohistochemistry for a-Smooth Muscle Actin and Cytokeratin-19, respectively. Angiogenesis was evaluated by ELISA and immunohistochemistry. Quantitative real-time PCR was used to determine the mRNA levels of liver Peroxisome Proliferator-Activated Receptor gamma (PPAR-gamma), Interleukin-6 (IL-6) and Tumor Necrosis-alpha (TNF alpha). BM-derived cells were tracked by double immunofluorescence. The spontaneous migration of mobilized HSCs towards injured liver and its cytokine secretion profile was determined in transwell culture systems. Either single-agent mobilization or the combination of agents significantly ameliorated hepatic damage by decreasing fibrosis and restoring the abnormal vascular network in the liver of mobilized mice compared to CCl4-only mice. The degree of fibrosis reduction was similar among all mobilized mice despite that G-CSF + Plerixafor yielded significantly higher numbers of circulating HSCs over other agents. The liver homing potential of variously mobilized HSCs differed among the agents. An extended G-CSF treatment provided the highest anti-fibrotic effect over all tested modalities, induced by the proliferation of hepatic stem cells and decreased hepatic inflammation. Plerixafor-mobilized HSCs, despite their reduced liver homing potential, reversed fibrosis mainly by increasing hepatic PPAR-gamma and VEGF expression. In all groups, BM-derived mature hepatocytes as well as liver-committed BM stem cells were detected only at low frequencies, further supporting the concept that alternative mechanisms rather than direct HSC effects regulate liver recovery. Overall, our data suggest that G-CSF, Plerixafor and G-CSF + Plerixafor act differentially during the wound healing process, ultimately providing a potent anti-fibrotic effect. (C) 2014 Elsevier Inc. All rights reserved.