Department of Ophthalmology, CHA Bundang Medical Center, CHA University, Seongnam, Gyeonggi-do, Republic of Korea.
Faculty of Life Sciences & Medicine, King's College London, London, SE5 9NU, UK.
Stem Cell Res Ther. 2019 Aug 9;10(1):246. doi: 10.1186/s13287-019-1348-0.
Graves' ophthalmopathy (GO) is a complication of Graves' disease (GD), in which orbital connective tissues become inflamed and increase in volume and orbital fibroblasts within the orbital fat and extraocular muscles differentiate into adipocytes in vitro when stimulated by hormones, several cytokines, and growth factors including TSH, IGF-1, IL-1, interferon γ, and platelet-derived growth factor. Human placental mesenchymal stem cells (hPMSCs) have immunomodulatory effects in disease pathogenesis. Although a number of studies have reported that hPMSCs can elicit therapeutic effects, these are not sufficient. Therefore, we constructed a GO animal model in order to find out the hPMSCs recovery effect.
We investigated their anti-adipogenic effects in in vitro cultures of orbital fibroblasts established from GO patients. Primary orbital fibroblasts were exposed to differentiation medium for 10 days. After being co-cultured with hPMSCs, the characteristics of orbital fibroblast were determined by Oil Red O stain and real-time PCR. Then, we explored the in vivo regulatory effects of hPMSCs in an experimental mouse model of GO. We developed the GO mouse model using immunization by leg muscle electroporation of pTriEx1.1Neo-hTSHR A-subunit plasmid. Human PMSC injection was performed into the left orbit. We also analyzed the effects of hPMSCs in the GO animal model.
We found that hPMSCs inhibited a lipid accumulation and activated factors, such as ADIPONECTIN, PPARγ, C/EBPα, and TGFβ2 genes in adipogenesis-induced primary orbital fibroblasts from GO patients. Moreover, hPMSCs were highly effective at ameliorating adipogenesis in the orbital tissue of the model.
These data indicate that hPMSCs recover pathogenic activation of orbital fibroblasts in animals undergoing experimental GO and confirm the feasibility of applying hPMSCs as a novel treatment for GO patients.
格雷夫斯眼病（GO）是格雷夫斯病（GD）的一种并发症，在这种疾病中，眼眶结缔组织发炎并肿大，眼眶脂肪和眼外肌中的成纤维细胞在激素、几种细胞因子和生长因子（包括 TSH、IGF-1、IL-1、干扰素 γ 和血小板衍生生长因子）的刺激下，在体外分化为脂肪细胞。人胎盘间充质干细胞（hPMSCs）在疾病发病机制中具有免疫调节作用。尽管许多研究报告称 hPMSCs 可以产生治疗效果，但这些效果还不够。因此，我们构建了 GO 动物模型，以了解 hPMSCs 的恢复效果。
我们研究了它们在从 GO 患者建立的眼眶成纤维细胞体外培养中的抗脂肪生成作用。原代眼眶成纤维细胞在分化培养基中培养 10 天。与 hPMSCs 共培养后，通过油红 O 染色和实时 PCR 确定眼眶成纤维细胞的特征。然后，我们在 GO 实验动物模型中探索了 hPMSCs 的体内调节作用。我们使用 pTriEx1.1Neo-hTSHR A 亚单位质粒经腿部肌肉电穿孔免疫来开发 GO 小鼠模型。将 hPMSC 注射到左侧眼眶。我们还分析了 hPMSCs 在 GO 动物模型中的作用。
我们发现 hPMSCs 抑制了 GO 患者原代眼眶成纤维细胞在脂肪生成诱导中的脂质积累，并激活了 ADIPONECTIN、PPARγ、C/EBPα 和 TGFβ2 等基因。此外，hPMSCs 非常有效地改善了模型眼眶组织中的脂肪生成。
这些数据表明，hPMSCs 可恢复实验性 GO 动物中眼眶成纤维细胞的致病激活，并证实了将 hPMSCs 作为 GO 患者的新型治疗方法的可行性。
