ClC-3 Deficiency Impairs the Neovascularization Capacity of Early Endothelial Progenitor Cells by Decreasing CXCR4/JAK-2 Signalling
ClC-3 Deficiency Impairs the Neovascularization Capacity of Early Endothelial Progenitor Cells by Decreasing CXCR4/JAK-2 Signalling
复制标题
ClC-3 缺乏通过减少 CXCR4-JAK-2 信号传导损害早期内皮祖细胞的新血管形成能力
DOI：
10.1016/j.cjca.2019.08.009
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发表时间：
2019-11-01
影响因子：
6.2
通讯作者：
Du, Yan-Hua
中科院分区：
文献类型：
--
作者：
Cheng, Li-Min;Li, Yue-Jiao;Du, Yan-Hua
背景：内皮祖细胞（Endothelial progenitor cell， EPC）治疗已被认为是缺血性疾病治疗的重大突破。然而，EPC功能调控的分子机制尚不清楚。方法：观察野生型和ClC-3敲除小鼠后肢缺血模型中EPCs的血管生成能力。结果：缺乏ClC-3的小鼠在后肢缺血后7天和14天的缺血肌肉血流恢复和新生血管形成均有所减少。此外，与野生型EPCs相比，接受ClC-3敲除EPCs的小鼠后肢血液再灌注明显受损，并伴有EPC归巢和保留减少。在体外，来自ClC-3敲除小鼠的EPCs表现出迁移、粘附和血管生成活性受损。与野生型相比，CXC趋化因子受体4 （CXCR4）在ClC-3敲除小鼠EPC中的表达显著降低。此外，CXCR4的下游信号通路Janus kinase 2 （JAK-2）的表达和磷酸化在ClC-3敲除的EPC中也降低，表明CXCR4/JAK-2信号通路因ClC-3缺乏而失调。与这一假设一致，野生型EPCs的迁移能力被CXCR4拮抗剂AMD3100或JAK-2抑制剂AG490削弱。更重要的是，过表达CXCR4可以挽救ClC-3敲除EPCs受损的迁移能力。结论：ClC-3在内皮祖细胞的血管生成能力和内皮祖细胞介导的缺血组织新生血管形成中起关键作用。CXCR4/JAK-2信号的紊乱可能导致ClC-3缺陷EPCs的功能损伤。因此，ClC-3可能是缺血性疾病中调节新生血管形成的潜在治疗靶点。
Background: Endothelial progenitor cell (EPC) therapy has been suggested as a major breakthrough in the treatment of ischemic diseases. However, the molecular mechanism that underlies EPC functional regulation is still unclear.Methods: We examined the angiogenic capacity of EPCs in a hindlimb ischemia model of wild-type and ClC-3 knockout mice.Results: Mice lacking of ClC-3 exhibited reduced blood flow recovery and neovascularization in ischemic muscles 7 and 14 days after hind limb ischemia. Moreover, compared with wild-type EPCs, the hindlimb blood reperfusion in mice receiving ClC-3 knockout EPCs was significantly impaired, accompanied by reduced EPC homing and retention. In vitro, EPCs derived from ClC-3 knockout mice displayed impaired migratory, adhesive, and angiogenic activity. CXC chemokine receptor 4 (CXCR4) expression was significantly reduced in EPC from ClC-3 knockout mice compared with wild-type. Moreover, the expression and phosphorylation of Janus kinase 2 (JAK-2), a downstream signalling of CXCR4, was also reduced in ClC-3 knockout EPC, indicating that CXCR4/JAK-2 signalling is dysregulated by ClC-3 deficiency. Consistent with this assumption, the migratory capacity of wild-type EPCs was attenuated by either CXCR4 antagonist AMD3100 or JAK-2 inhibitor AG490. More importantly, the impaired migratory capacity of ClC-3 knockout EPCs was rescued by overexpression of CXCR4.Conclusions: ClC-3 plays a critical role in the angiogenic capacity of EPCs and EPC-mediated neovascularization of ischemic tissues. Disturbance of CXCR4/JAK-2 signalling may contribute to the functional impairment of ClC-3 deficient EPCs. Thus, ClC-3 may be a potential therapeutic target for modulating neovascularization in ischemic diseases.