Hypoxia preconditioning promotes cardiac stem cell survival and cardiogenic differentiation in vitro involving activation of the HIF-1α/apelin/APJ axis.
Hypoxia preconditioning promotes cardiac stem cell survival and cardiogenic differentiation in vitro involving activation of the HIF-1α/apelin/APJ axis.
复制标题
缺氧预处理促进心脏干细胞存活和体外心源性分化，涉及 HIF-1 α/apelin/APJ 轴的激活
DOI：
10.1186/s13287-017-0673-4
复制
发表时间：
2017-09-29
影响因子：
7.5
通讯作者：
Wang T
中科院分区：
文献类型：
--
作者：
Hou J;Wang L;Long H;Wu H;Wu Q;Zhong T;Chen X;Zhou C;Guo T;Wang T
心脏干细胞（CSCs）移植被认为是心血管疾病的一种理想治疗方法。然而，这些细胞在局部梗死部位的存活率低且分化效率差，降低了其治疗效果。在本研究中，我们在体外研究了缺氧预处理（HP）对心脏干细胞存活和向心肌细胞分化的影响，并探讨了相关机制。
心脏干细胞取自斯普拉格 - 道利大鼠，将第3代细胞在体外培养并暴露于缺氧环境（1% O₂）。分别通过MTS法和流式细胞术评估细胞存活率和凋亡情况。在细胞经历缺氧预处理后，再用5 - 氮杂胞苷诱导24小时以促使其向心肌细胞分化。在整个过程中，常氧（20% O₂）作为阴性对照。在诱导2周后评估向心肌细胞分化的情况。在缺氧预处理后以及分化过程中检测相关分子。转染抗缺氧诱导因子 - 1α（HIF - 1α）小干扰RNA（siRNA）、抗apelin siRNA和抗血管紧张素受体AT1相关的假定受体蛋白（APJ）siRNA以阻断它们的表达，并检测相关下游分子。
与常氧组相比，缺氧组在12小时和24小时时间点生长更快（p < 0.01）。细胞在24小时时间点呈现出最高的增殖率（p < 0.01）。缺氧暴露24小时后细胞凋亡率显著下降（p < 0.01）。缺氧预处理后HIF - 1α、apelin和APJ的表达水平均升高。诱导2周后，缺氧预处理组中apelin、α - 肌动蛋白（α - SA）和心肌肌钙蛋白T（cTnT）阳性细胞的百分比大幅增加。在7天和14天，α - SA和cTnT的蛋白水平也显著升高（p < 0.01）。在向心肌细胞分化过程中的不同时间点，HIF - 1α、apelin和APJ均升高（p < 0.01）。通过siRNAs敲低HIF - 1α、apelin或APJ导致α - SA和cTnT显著减少。阻断HIF - 1α导致apelin和APJ显著降低（p < 0.01）。抑制apelin后，apelin和APJ的表达水平下降（p < 0.01）。
缺氧预处理可有效促进体外心脏干细胞的存活和向心肌细胞分化，这一过程涉及HIF - 1α/apelin/APJ轴的激活。本研究为探索提高心脏干细胞移植效率的新策略提供了新的视角。
Cardiac stem cells (CSCs) transplantation has been regarded as an optimal therapeutic approach for cardiovascular disease. However, inferior survival and low differentiation efficiency of these cells in the local infarct site reduce their therapeutic efficacy. In this study, we investigated the influence of hypoxia preconditioning (HP) on CSCs survival and cardiogenic differentiation in vitro and explored the relevant mechanism. CSCs were obtained from Sprague–Dawley rats and cells of the third passage were cultured in vitro and exposed to hypoxia (1% O2). Cells survival and apoptosis were evaluated by MTS assay and flow cytometry respectively. Cardiogenic differentiation was induced by using 5-azacytidine for another 24 h after the cells experienced HP. Normoxia (20% O2) was used as a negative control during the whole process. Cardiogenic differentiation was assessed 2 weeks after the induction. Relevant molecules were examined after HP and during the differentiation process. Anti-hypoxia-inducible factor-1α (HIF-1α) small interfering RNA (siRNA), anti-apelin siRNA, and anti-putative receptor protein related to the angiotensin receptor AT1 (APJ) siRNA were transfected in order to block their expression, and relevant downstream molecules were detected. Compared with the normoxia group, the hypoxia group presented more rapid growth at time points of 12 and 24 h (p < 0.01). Cells exhibited the highest proliferation rate at the time point of 24 h (p < 0.01). The cell apoptosis rate significantly declined after 24 h of hypoxia exposure (p < 0.01). Expression levels of HIF-1α, apelin, and APJ were all enhanced after HP. The percentage of apelin, α-SA, and cTnT positive cells was greatly increased in the HP group after 2 weeks of induction. The protein level of α-SA and cTnT was also significantly elevated at 7 and 14 days (p < 0.01). HIF-1α, apelin, and APJ were all increased at different time points during the cardiogenic differentiation process (p < 0.01). Knockdown of HIF-1α, apelin or APJ by siRNAs resulted in a significant reduction of α-SA and cTnT. HIF-1α blockage caused a remarkable decrease of apelin and APJ (p < 0.01). Expression levels of apelin and APJ were depressed after the inhibition of apelin (p < 0.01). HP could effectively promote CSCs survival and cardiogenic differentiation in vitro, and this procedure involved activation of the HIF-1α/apelin/APJ axis. This study provided a new perspective for exploring novel strategies to enhance CSCs transplantation efficiency.
登录
查看更多内容
影响因子：
2.6
作者：
Chen J;Kang JG;Keyvanfar K;Young NS;Hwang PM
通讯作者：
Hwang PM
心脏干细胞移植通过ANG II/AT1R/TGF-β1信号通路增强心肌梗死大鼠模型中连接蛋白43的表达
影响因子：
3.6
作者：
Hou, Jingying;Yan, Ping;Wang, Tong
通讯作者：
Wang, Tong
影响因子：
4.7
作者：
Mottaghi, S.;Larijani, B.;Sharifi, Ali M.
通讯作者：
Sharifi, Ali M.
影响因子：
4
作者：
Cerrada, Inmaculada;Ruiz-Sauri, Amparo;Sepulveda, Pilar
通讯作者：
Sepulveda, Pilar
影响因子：
64.8
作者：
Kimura, Wataru;Xiao, Feng;Sadek, Hesham A.
通讯作者：
Sadek, Hesham A.