Isildar Basak, Ozkan Serbay, Sahin Hakan, Ercin Merve, Gezginci-Oktayoglu Selda, Koyuturk Meral
Department of Histology and Embryology, Cerrahpasa Faculty of Medicine, Istanbul University-Cerrahpasa, Istanbul, Turkey; Balikesir University, Faculty of Medicine, Histology and Embryology Department, Balikesir, Turkey.
Department of Histology and Embryology, Cerrahpasa Faculty of Medicine, Istanbul University-Cerrahpasa, Istanbul, Turkey; Izmir Katip Celebi University, Faculty of Medicine, Histology and Embryology Department, Izmir, Turkey.
Int Immunopharmacol. 2024 Mar 10;129:111662. doi: 10.1016/j.intimp.2024.111662. Epub 2024 Feb 9.
This study aimed to examine the effects of the secretome released by human umbilical cord-mesenchymal stem cells (MSC) as a result of preconditioning with deferoxamine (DFX), a hypoxia mimetic agent, on type 1 diabetes (T1D), by comparing it with the secretome produced by untreated MSCs. Initially, the levels of total protein, IL4, IL10, IL17, and IFNγ in the conditioned medium (CM) obtained from MSCs subjected to preconditioning with 150 µM DFX (DFX-CM) were analyzed in comparison to CM derived from untreated MSCs (N-CM). Subsequently, the CMs were administered to rats with T1D within a specific treatment plan. Following the sacrification, immunomodulation was evaluated by measuring serum cytokine levels and assessing the regulatory T cell (Treg) ratio in spleen mononuclear cells. Additionally, β-cell mass was determined in the islets by immunohistochemical labeling of NK6 Homeobox 1 (Nkx6.1), Pancreatic duodenal homeobox-1 (Pdx1), and insulin antibodies in pancreatic sections. In vitro findings indicated that the secretome levels of MSCs were enhanced by preconditioning with DFX. In vivo, the use of DFX-CM significantly increased the Treg population, and accordingly, the level of inflammatory cytokines decreased. In β-cell marker labeling, D + DFX-CM showed significantly increased PDX1 and insulin immunoreactivity. In conclusion, while the factors released by MSCs without external stimulation had limited therapeutic effects, substantial improvements in immunomodulation and β-cell regeneration were seen with DFX-preconditioned cell-derived CM.
本研究旨在通过将人脐带间充质干细胞（MSC）经去铁胺（DFX，一种缺氧模拟剂）预处理后释放的分泌组与未处理的MSC产生的分泌组进行比较，来研究其对1型糖尿病（T1D）的影响。最初，分析了从用150µM DFX预处理的MSC获得的条件培养基（CM）（DFX-CM）中总蛋白、IL4、IL10、IL17和IFNγ的水平，并与从未处理的MSC获得的CM（N-CM）进行比较。随后，按照特定治疗方案将这些CM施用于T1D大鼠。处死大鼠后，通过测量血清细胞因子水平和评估脾单核细胞中的调节性T细胞（Treg）比例来评估免疫调节。此外，通过胰腺切片中NK6同源盒1（Nkx6.1）、胰腺十二指肠同源盒-1（Pdx1）和胰岛素抗体的免疫组织化学标记来测定胰岛中的β细胞量。体外研究结果表明，DFX预处理可提高MSC的分泌组水平。在体内，使用DFX-CM可显著增加Treg群体，相应地，炎性细胞因子水平降低。在β细胞标志物标记中，D + DFX-CM显示PDX1和胰岛素免疫反应性显著增加。总之，虽然未经外部刺激的MSC释放的因子具有有限的治疗效果，但DFX预处理的细胞来源的CM在免疫调节和β细胞再生方面有显著改善。
