Ang II-AT2R increases mesenchymal stem cell migration by signaling through the FAK and RhoA/Cdc42 pathways in vitro.
Ang II-AT2R increases mesenchymal stem cell migration by signaling through the FAK and RhoA/Cdc42 pathways in vitro.
复制标题
Ang II-AT2R 通过体外 FAK 和 RhoA/Cdc42 通路信号传导增加间充质干细胞迁移
DOI：
10.1186/s13287-017-0617-z
复制
发表时间：
2017-07-12
影响因子：
7.5
通讯作者：
Qiu HB
中科院分区：
文献类型：
--
作者：
Xu XP;He HL;Hu SL;Han JB;Huang LL;Xu JY;Xie JF;Liu AR;Yang Y;Qiu HB
背景间充质干细胞（MSC）通过血流迁移到损伤部位，并可能被炎症因子吸引。据报道，作为一种促炎介质，血管紧张素 II (Ang II) 通过体外 Ang II 受体的信号传导增强各种细胞类型的迁移。然而，很少有研究关注Ang II对MSC迁移的影响及其潜在机制。方法采用不同浓度的Ang II、AT1R拮抗剂（氯沙坦）和/或AT2R拮抗剂（PD-123319）治疗后，使用伤口愈合和博伊登室迁移测定法测量人骨髓MSC的迁移。为了排除增殖对 MSC 迁移的影响，我们测量了用相同浓度的 Ang II 刺激后 MSC 的增殖情况。此外，我们使用粘着斑激酶 (FAK) 抑制剂 PF-573228、RhoA 抑制剂 C3 转移酶、Rac1 抑制剂 NSC23766 或 Cdc42 抑制剂 ML141 来研究细胞粘附蛋白和 Rho-GTPase 蛋白家族（RhoA、Rac1 和 Cdc42）在 Ang II 介导的 MSC 迁移中的作用。通过蛋白质印迹分析检测细胞粘附蛋白（FAK、Talin 和 Vinculin）。通过G-LISA评估Rho-GTPase家族蛋白活性，免疫荧光检测反映肌动蛋白细胞骨架组织的F-actin水平。结果人骨髓MSCs持续表达AT1R和AT2R。此外，Ang II 以 AT2R 依赖性方式增加 MSC 迁移。值得注意的是，Ang II 增强的迁移不是由 Ang II 介导的细胞增殖介导的。有趣的是，Ang II 增强的迁移是由 FAK 激活介导的，这对于焦点接触的形成至关重要，Talin 和 Vinculin 表达增加就证明了这一点。此外，FAK 激活 RhoA 和 Cdc42，增加细胞骨架组织，从而促进细胞收缩。此外，PD-123319 阻止了响应 Ang II 的 FAK、Talin 和 Vinculin 激活以及 F-肌动蛋白重组，但洛沙坦则不然，表明 FAK 激活和 F-肌动蛋白重组是 AT2R 的下游。结论这些数据表明 Ang II-AT2R 通过 FAK 和 FAK 信号传导来调节人骨髓 MSC 迁移。 RhoA/Cdc42 pathways.这项研究提供了对 MSC 归巢至损伤部位的机制的见解，并将有助于合理设计靶向疗法以改善 MSC 植入。
BackgroundMesenchymal stem cells (MSCs) migrate via the bloodstream to sites of injury and are possibly attracted by inflammatory factors. As a proinflammatory mediator, angiotensin II (Ang II) reportedly enhances the migration of various cell types by signaling via the Ang II receptor in vitro. However, few studies have focused on the effects of Ang II on MSC migration and the underlying mechanisms.MethodsHuman bone marrow MSCs migration was measured using wound healing and Boyden chamber migration assays after treatments with different concentrations of Ang II, an AT1R antagonist (Losartan), and/or an AT2R antagonist (PD-123319). To exclude the effect of proliferation on MSC migration, we measured MSC proliferation after stimulation with the same concentration of Ang II. Additionally, we employed the focal adhesion kinase (FAK) inhibitor PF-573228, RhoA inhibitor C3 transferase, Rac1 inhibitor NSC23766, or Cdc42 inhibitor ML141 to investigate the role of cell adhesion proteins and the Rho-GTPase protein family (RhoA, Rac1, and Cdc42) in Ang II-mediated MSC migration. Cell adhesion proteins (FAK, Talin, and Vinculin) were detected by western blot analysis. The Rho-GTPase family protein activities were assessed by G-LISA and F-actin levels, which reflect actin cytoskeletal organization, were detected by using immunofluorescence.ResultsHuman bone marrow MSCs constitutively expressed AT1R and AT2R. Additionally, Ang II increased MSC migration in an AT2R-dependent manner. Notably, Ang II-enhanced migration was not mediated by Ang II-mediated cell proliferation. Interestingly, Ang II-enhanced migration was mediated by FAK activation, which was critical for the formation of focal contacts, as evidenced by increased Talin and Vinculin expression. Moreover, RhoA and Cdc42 were activated by FAK to increase cytoskeletal organization, thus promoting cell contraction. Furthermore, FAK, Talin, and Vinculin activation and F-actin reorganization in response to Ang II were prevented by PD-123319 but not Losartan, indicating that FAK activation and F-actin reorganization were downstream of AT2R.ConclusionsThese data indicate that Ang II-AT2R regulates human bone marrow MSC migration by signaling through the FAK and RhoA/Cdc42 pathways. This study provides insights into the mechanisms by which MSCs home to injury sites and will enable the rational design of targeted therapies to improve MSC engraftment.
登录
查看更多内容
DOI：
10.1161/hypertensionaha.112.203679
发表时间：
2013-01
期刊：
Hypertension (Dallas, Tex. : 1979)
影响因子：
--
作者：
Herrera M;Sparks MA;Alfonso-Pecchio AR;Harrison-Bernard LM;Coffman TM
通讯作者：
Coffman TM
影响因子：
2.5
作者：
Hellmann, MA;Panet, H;Offen, D
通讯作者：
Offen, D
影响因子：
8.1
作者：
Li, Li;Jiang, Jianxin
通讯作者：
Jiang, Jianxin
人间充质干细胞对活细菌损伤的离体人肺的治疗作用
DOI：
10.1164/rccm.201206-0990oc
发表时间：
2013-04-01
影响因子：
24.7
作者：
Lee, Jae W.;Krasnodembskaya, Anna;Matthay, Michael A.
通讯作者：
Matthay, Michael A.
影响因子：
5.6
作者：
Cai, Shi-Xia;Liu, Ai-Ran;Qiu, Hai-Bo
通讯作者：
Qiu, Hai-Bo