Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 31-007 Kraków, Poland.
Basic Science Department, College of Agriculture, University of Duhok, Zakho Street 38, 1006 AJ Duhok, Kurdistan Region, Iraq.
Mediators Inflamm. 2019 Jul 16;2019:1868170. doi: 10.1155/2019/1868170. eCollection 2019.
Myeloid angiogenic cells (MAC) derive from hematopoietic stem/progenitor cells (HSPCs) that are mobilized from the bone marrow. They home to sites of neovascularization and contribute to angiogenesis by production of paracrine factors. The number and function of proangiogenic cells are impaired in patients with diabetes or cardiovascular diseases. Both conditions can be accompanied by decreased levels of heme oxygenase-1 (HMOX1), cytoprotective, heme-degrading enzyme. Our study is aimed at investigating whether precursors of myeloid angiogenic cells (PACs) treated with known pharmaceuticals would produce media with better proangiogenic activity and if such media can be used to stimulate blood vessel growth . We used G-CSF-mobilized CD34 HSPCs, FACS-sorted from healthy donor peripheral blood mononuclear cells (PBMCs). Sorted cells were predominantly CD133. CD34 cells after six days in culture were stimulated with atorvastatin (AT), acetylsalicylic acid (ASA), sulforaphane (SR), resveratrol (RV), or metformin (Met) for 48 h. Conditioned media from such cells were then used to stimulate human aortic endothelial cells (HAoECs) to enhance tube-like structure formation in a Matrigel assay. The only stimulant that enhanced PAC paracrine angiogenic activity was atorvastatin, which also had ability to stabilize endothelial tubes . On the other hand, the only one that induced heme oxygenase-1 expression was sulforaphane, a known activator of a HMOX1 inducer-NRF2. None of the stimulants changed significantly the levels of 30 cytokines and growth factors tested with the multiplex test. Then, we used atorvastatin-stimulated cells or conditioned media from them in the Matrigel plug angiogenic assay. Neither AT alone in control media nor conditioned media nor AT-stimulated cells affected numbers of endothelial cells in the plug or plug's vascularization. Concluding, high concentrations of atorvastatin stabilize tubes and enhance the paracrine angiogenic activity of human PAC cells . However, the effect was not observed . Therefore, the use of conditioned media from atorvastatin-treated PAC is not a promising therapeutic strategy to enhance angiogenesis.
骨髓源性血管生成细胞 (MAC) 来源于造血干细胞/祖细胞 (HSPC)，这些细胞从骨髓中动员而来。它们归巢到新血管生成部位，并通过产生旁分泌因子促进血管生成。患有糖尿病或心血管疾病的患者，其促血管生成细胞的数量和功能受损。这两种情况都可能伴有血红素加氧酶-1 (HMOX1) 水平降低，血红素加氧酶-1 是一种细胞保护性的、血红素降解酶。我们的研究旨在调查用已知药物处理的骨髓源性血管生成细胞 (PAC) 前体是否会产生具有更好促血管生成活性的培养基，如果可以使用这种培养基来刺激血管生长。我们使用 G-CSF 动员的 CD34 HSPC，从健康供体外周血单个核细胞 (PBMC) 中通过 FACS 分选。分选细胞主要是 CD133。CD34 细胞在培养 6 天后用阿托伐他汀 (AT)、乙酰水杨酸 (ASA)、萝卜硫素 (SR)、白藜芦醇 (RV) 或二甲双胍 (Met) 刺激 48 小时。然后用这种细胞的条件培养基刺激人主动脉内皮细胞 (HAoEC)，以增强在 Matrigel 测定中的管状结构形成。唯一增强 PAC 旁分泌血管生成活性的刺激物是阿托伐他汀，它还具有稳定内皮管的能力。另一方面，唯一诱导血红素加氧酶-1 表达的是萝卜硫素，它是一种已知的血红素加氧酶-1 诱导物-NRF2 激活剂。在多重测试中，没有一种刺激物显著改变 30 种细胞因子和生长因子的水平。然后，我们在 Matrigel 塞血管生成测定中使用阿托伐他汀刺激的细胞或它们的条件培养基。单独在对照培养基中的 AT 或条件培养基或 AT 刺激的细胞均不影响塞中的内皮细胞数量或塞的血管化。总之，阿托伐他汀的高浓度可稳定管腔并增强人 PAC 细胞的旁分泌血管生成活性。然而，这种作用并未观察到。因此，使用阿托伐他汀处理后的 PAC 的条件培养基不是增强血管生成的有前途的治疗策略。
