Department of Surgery, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
European Center of Angioscience (ECAS), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
Stem Cell Res Ther. 2023 Feb 9;14(1):24. doi: 10.1186/s13287-022-03228-w.
Hypoxia in ischemic disease impairs Ca homeostasis and may promote angiogenesis. The therapeutic efficacy of mesenchymal stromal cells (MSCs) in peripheral arterial occlusive disease is well established, yet its influence on cellular Ca homeostasis remains to be elucidated. We addressed the influence of ATP-binding cassette subfamily B member 5 positive mesenchymal stromal cells (ABCB5 MSCs) on Ca homeostasis in hypoxic human umbilical vein endothelial cells (HUVECs) in vitro and in vivo.
Hypoxia was induced in HUVECs by Cobalt (II) chloride (CoCl) or Deferoxamine (DFO). Dynamic changes in the cytosolic- and endoplasmic reticulum (ER) Ca and changes in reactive oxygen species were assessed by appropriate fluorescence-based sensors. Metabolic activity, cell migration, and tube formation were assessed by standard assays. Acute-on-chronic ischemia in Apolipoprotein E knock-out (ApoE) mice was performed by double ligation of the right femoral artery (DFLA). ABCB5 MSC cells were injected into the ischemic limb. Functional recovery after DFLA and histology of gastrocnemius and aorta were assessed.
Hypoxia-induced impairment of cytosolic and ER Ca were restored by ABCB5 MSCs or their conditioned medium. Similar was found for changes in intracellular ROS production, metabolic activity, migratory ability and tube formation. The restoration was paralleled by an increased expression of the Ca transporter Sarco-/endoplasmic reticulum ATPase 2a (SERCA2a) and the phosphorylation of Phospholamban (PLN). In acute-on-chronic ischemia, ABCB5 MSCs treated mice showed a higher microvascular density, increased SERCA2a expression and PLN phosphorylation relative to untreated controls.
ABCB5 MSCs therapy can restore cellular Ca homeostasis, which may beneficially affect the angiogenic function of endothelial cells under hypoxia in vitro and in vivo.
缺血性疾病中的缺氧会损害钙稳态，并可能促进血管生成。间充质基质细胞（MSCs）在周围动脉闭塞性疾病中的治疗效果已得到充分证实，但它对细胞钙稳态的影响仍有待阐明。我们研究了 ATP 结合盒亚家族 B 成员 5 阳性间充质基质细胞（ABCB5 MSC）对体外和体内缺氧人脐静脉内皮细胞（HUVEC）钙稳态的影响。
通过氯化钴（CoCl）或去铁胺（DFO）诱导 HUVEC 缺氧。通过适当的荧光传感器评估细胞质和内质网（ER）钙的动态变化以及活性氧的变化。通过标准测定评估代谢活性、细胞迁移和管状形成。通过双重结扎右股动脉（DFLA）在载脂蛋白 E 敲除（ApoE）小鼠中进行急性慢性缺血。将 ABCB5 MSC 细胞注入缺血肢体。评估 DFLA 后的功能恢复和比目鱼肌和主动脉的组织学。
ABCB5 MSC 或其条件培养基可恢复缺氧诱导的细胞质和 ER 钙损伤。细胞内 ROS 产生、代谢活性、迁移能力和管状形成的变化也是如此。这种恢复与钙转运体肌浆/内质网 ATP 酶 2a（SERCA2a）的表达增加和磷蛋白磷酸化（PLN）平行。在急性慢性缺血中，与未治疗的对照组相比，接受 ABCB5 MSC 治疗的小鼠显示出更高的微血管密度、SERCA2a 表达增加和 PLN 磷酸化。
ABCB5 MSC 治疗可以恢复细胞钙稳态，这可能有益于体外和体内缺氧条件下内皮细胞的血管生成功能。
