Interruption of CD40 Pathway Improves Efficacy of Transplanted Endothelial Progenitor Cells in Monocrotaline Induced Pulmonary Arterial Hypertension
Interruption of CD40 Pathway Improves Efficacy of Transplanted Endothelial Progenitor Cells in Monocrotaline Induced Pulmonary Arterial Hypertension
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DOI：
10.1159/000430130
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发表时间：
2015-05-01
影响因子：
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通讯作者：
Wang, XingXiang
中科院分区：
文献类型：
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作者：
Pan, YanYun;Wang, Shuai;Wang, XingXiang
背景/目的：内皮祖细胞移植对肺动脉高压(PAH)有治疗作用。同时，祖细胞的招募具有潜在的炎症效应，并夸大了血管损伤。CD40通路被认为是血管炎症事件的主要参与者。在本研究中，我们研究了CD40通路在调节早期生长的EPC功能中的作用，并寻求PAH细胞治疗的改进。方法：从大鼠骨髓中分离培养内皮祖细胞，培养7d。经可溶性CD40配体(SCD4OL)处理24小时后，检测EPC的迁移、黏附、增殖、旁分泌和血管生成功能。采用野百合碱(MCT)皮下注射建立大鼠PAH模型。分别于注射MCT后第7、14、21天经尾静脉注入对照EPC或慢病毒载体(Lv)-shRNA-CD40 EPC。在第28天评价治疗效果。结果：sCD4OL剂量依赖性地损害EPC的迁移、黏附、增殖和血管生成功能。而sCD4OL对可溶性细胞间黏附分子-1、血管内皮生长因子和白介素6的旁分泌作用呈剂量依赖性增强。对照EPC来源的条件培养液对体外血管生成有保护作用，而sCD40L处理的内皮细胞则表现出不利的作用。注射MCT后，大鼠血清sCD4OL水平逐渐升高。除体外结果外，两种EPC治疗的益处都很明显，即使在第21天服用也是如此。对照内皮祖细胞的益处随着时间的推移逐渐消失，但Lv-shRNA-CD40内皮祖细胞的益处更有效和持久，其特征是改善了大鼠的血流动力学，逆转了血管重塑。此外，Lv-shRNA-CD40内皮祖细胞能更好地整合到内皮细胞中，而不是整合到外膜和中膜中。结论：sCD4OL抑制内皮祖细胞的保护作用。传统的EPC治疗局限于PAH，而阻断移植细胞的CD40通路可明显提高治疗效果。版权所有(C)2015年S·卡格尔股份公司，巴塞尔
Background/Aims: Transplantation of endothelial progenitor cells (EPCs) plays a therapeutic role in pulmonary arterial hypertension (PAH). Meanwhile, recruitment of progenitors has potential inflammatory effects and exaggerates vascular injury. CD40 pathway is identified as a major player in vascular inflammatory events. In this study, we investigated the role of CD40 pathway in regulating early outgrowth EPC functions, and searched for improvements in PAH cell therapy. Methods: EPCs were isolated from rat bone marrow and cultured for 7 days. After treatment with soluble CD40 ligand (sCD4OL) for 24 hours, EPC migration, adhesion, proliferation, paracrine and vasculogenesis functions were tested. Rat PAH model was founded by subcutaneous injection of monocrotaline (MCT). Control EPCs or lentivirus vectors (Lv)-shRNA-CD40 EPCs were infused via tail vein at day 7, 14, and 21 after MCT injection. Therapeutic effects were evaluated at day 28. Results: sCD4OL dose-dependently impaired EPC migration, adhesion, proliferation, and vasculogenesis functions. However, paracrine effects of soluble intercellular adhesion molecule-1, vascular endothelial growth factor and interleukin-6 were dose-dependently improved by sCD4OL. Control EPC-derived conditioned medium protected endothelial cell in vitro vasculogenesis, while sCD40L-pretreated ones showed detrimental effects. After MCT injection, sCD4OL levels in rat serum increased gradually. Other than in vitro results, benefits of both two EPC treatments were obvious, even taken at day 21. Benefits of control EPCs wore off over time, but those of Lv-shRNA-CD40 EPCs were more effective and enduring, as characterized by both ameliorated rat hemodynamic and reversed vascular remodeling. Furthermore, Lv-shRNA-CD40 EPCs integrated into endothelium better, rather than into adventitia and media. Conclusion: sCD4OL impaired protective effects of EPCs. Traditional EPC treatments were limited in PAH, while interruption of CD40 pathway of transplanted cells could apparently improve the therapeutic efficacy. Copyright (C) 2015 S Karger AG, Basel