Fadaghi Shohreh, Mahmoodi Merat, Derakhshani Ali, Sedghy Farnaz, Ranjkesh Mahdi, Behzadi Ahmadreza
Department of Immunology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
Research Center for Hydatid Disease in Iran, Kerman University of Medical Sciences, Kerman, Iran.
Heliyon. 2024 Aug 2;10(16):e35667. doi: 10.1016/j.heliyon.2024.e35667. eCollection 2024 Aug 30.
In this study, mesenchymal stem cells (MSCs) were primed with Tetrandrine (TET) having anti-inflammatory and immunomodulatory effects to examine the effects of this molecule on the antioxidative potential of MSCs as well as their modulatory effects on activated peripheral blood mononuclear cells (PBMCs). The viability of primed MSCs was detected using MTT assay and Trypan blue staining. Moreover, flow cytometry technique was applied to evaluate cell cycle distribution and immunophenotype of MSCs. The production of superoxide dismutase 3 (SOD3), malondialdehyde (MDA), kynurenine, TGF-β, and IFN-γ were also measured by spectrophotometry to assess the alteration of antioxidative and immunomodulatory potential of MSCs. Then, TET-primed MSCs were cocultured with PBMCs. The MTT assay was used to measure the proliferation of PBMCs. Cell cycle progression of PBMCs and frequency of regulatory T cells were evaluated using Flow cytometry. ELISA assay was also applied to determine the concentrations of TGF-β and IFN-γ after coculturing. According to our data, TET enhanced the secretion of SOD3 and kynurenine from MSCs, while the production of IFN-γ was reduced. No changes were observed in the viability, proliferation, and immunophenotype of MSCs after priming with TET. Moreover, the proliferation and frequency of PBMCs in the S and G2/M phases of cell cycle reduced after co-culturing with TET-primed MSCs. The concentration of TGF-β was increased in the supernatant of PBMCs, but the level of IFN-γ was reduced. Our data suggested this priming method as a novel strategy for increasing the antioxidative and immunomodulatory activity of MSCs.
在本研究中，用具有抗炎和免疫调节作用的粉防己碱（TET）预处理间充质干细胞（MSC），以研究该分子对MSC抗氧化潜能的影响及其对活化外周血单个核细胞（PBMC）的调节作用。使用MTT法和台盼蓝染色检测预处理后MSC的活力。此外，应用流式细胞术技术评估MSC的细胞周期分布和免疫表型。还通过分光光度法测量超氧化物歧化酶3（SOD3）、丙二醛（MDA）、犬尿氨酸、转化生长因子-β（TGF-β）和干扰素-γ（IFN-γ）的产生，以评估MSC抗氧化和免疫调节潜能的变化。然后，将经TET预处理的MSC与PBMC共培养。使用MTT法测量PBMC的增殖。使用流式细胞术评估PBMC的细胞周期进程和调节性T细胞的频率。共培养后还应用酶联免疫吸附测定（ELISA）法测定TGF-β和IFN-γ的浓度。根据我们的数据，TET增强了MSC中SOD3和犬尿氨酸的分泌，同时降低了IFN-γ的产生。用TET预处理后，MSC的活力、增殖和免疫表型未观察到变化。此外，与经TET预处理的MSC共培养后，PBMC在细胞周期S期和G2/M期的增殖和频率降低。PBMC上清液中TGF-β的浓度增加，但IFN-γ的水平降低。我们的数据表明这种预处理方法是提高MSC抗氧化和免疫调节活性的一种新策略。
