Reyner Claudia L, Winter Randolph L, Maneval Kara L, Boone Lindsey H, Wooldridge Anne A
Am J Vet Res. 2021 Apr;82(4):318-325. doi: 10.2460/ajvr.82.4.318.
To investigate the effects of recombinant equine IL-1β on function of equine endothelial colony-forming cells (ECFCs) in vitro.
ECFCs derived from peripheral blood samples of 3 healthy adult geldings.
Function testing was performed to assess in vitro wound healing, tubule formation, cell adhesion, and uptake of 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate-labeled acetylated low-density lipoprotein (DiI-Ac-LDL) by cultured ECFCs. Cell proliferation was determined by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay. Effects on function test results of different concentrations and exposure times of recombinant equine IL-1β were assessed.
Challenge of cultured ECFCs with IL-1β for 48 hours inhibited tubule formation. Continuous challenge (54 hours) with IL-1β in the wound healing assay reduced gap closure. The IL-1β exposure did not significantly affect ECFC adhesion, DiI-Ac-LDL uptake, or ECFC proliferation.
These results suggested a role for IL-1β in the inhibition of ECFC function in vitro. Functional changes in ECFCs following challenge with IL-1β did not appear to be due to changes in cell proliferative capacity. These findings have implications for designing microenvironments for and optimizing therapeutic effects of ECFCs used to treat ischemic diseases in horses.
研究重组马白细胞介素-1β（IL-1β）对马内皮祖细胞（ECFCs）体外功能的影响。
来自3匹健康成年去势公马外周血样本的ECFCs。
进行功能测试以评估培养的ECFCs的体外伤口愈合、小管形成、细胞黏附以及对1,1'-二油酰基-3,3,3',3'-四甲基吲哚羰花青高氯酸盐标记的乙酰化低密度脂蛋白（DiI-Ac-LDL）的摄取。通过2,3-双-(2-甲氧基-4-硝基-5-磺基苯基)-2H-四唑-5-羧基苯胺检测法测定细胞增殖。评估不同浓度和暴露时间的重组马IL-1β对功能测试结果的影响。
用IL-1β刺激培养的ECFCs 48小时可抑制小管形成。在伤口愈合试验中用IL-1β持续刺激（54小时）可减少缺口闭合。IL-1β暴露对ECFC黏附、DiI-Ac-LDL摄取或ECFC增殖无显著影响。
这些结果表明IL-1β在体外抑制ECFC功能中起作用。用IL-1β刺激后ECFCs的功能变化似乎并非由于细胞增殖能力的改变。这些发现对于设计用于治疗马缺血性疾病的ECFCs的微环境和优化其治疗效果具有重要意义。
