摘要
背景:强直性脊柱炎是一种伴有高炎性状态的自身免疫性疾病,但其具体发病机制不明确,目前尚缺乏有效的治疗方案。目的:体外探讨强直性脊柱炎患者来源骨髓间充质干细胞调控巨噬细胞的功能及正常人骨髓间充质干细胞在强直性脊柱炎治疗中的潜在作用。方法:提取25例强直性脊柱炎患者及21例健康志愿者的骨髓间充质干细胞进行体外培养扩增,以第4代细胞进行实验。体外诱导THP-1细胞系分化成为巨噬细胞。运用流式细胞术鉴定骨髓间充质干细胞及巨噬细胞的表面标志。ELISA检测健康志愿者/强直性脊柱炎患者骨髓间充质干细胞与巨噬细胞共培养体系上清中肿瘤坏死因子α及TSG-6蛋白水平,PCR检测巨噬细胞及骨髓间充质干细胞相关因子基因表达水平。结果与结论:(1)两种骨髓间充质干细胞均呈现典型间充质干细胞表型,巨噬细胞表型标志CD68为阳性。(2)强直性脊柱炎患者骨髓间充质干细胞与巨噬细胞共培养组中巨噬细胞分泌的肿瘤坏死因子α蛋白水平及肿瘤坏死因子α转录水平均高于健康志愿者骨髓间充质干细胞(P<0.05)。(3)强直性脊柱炎患者骨髓间充质干细胞组TSG-6基因的转录水平及蛋白分泌水平均低于健康志愿者骨髓间充质干细胞组(P<0.05)。结果提示:(1)强直性脊柱炎患者骨髓间充质干细胞抑制巨噬细胞肿瘤坏死因子α分泌的功能减弱,可能是导致强直性脊柱炎免疫异常的重要原因。(2)正常人骨髓间充质干细胞可分泌足量的TSG-6抑制强直性脊柱炎患者巨噬细胞的活化,降低其肿瘤坏死因子α的分泌而发挥治疗作用。
BACKGROUND： Ankylosing spondylitis is an autoimmune disease at high inflammatory state, and its pathogenesis is still unclear. Besides, there is a lack of entirely satisfactory curative strategies. OBJECTIVE： To explore the immunoregulation capability of bone marrow mesenchymal stem cells from ankylosing spondylitis patients on macrophages and the potential therapeutic use of bone marrow mesenchymal stem cells from healthy donors on ankylosing spondylitis. METHODS： Bone marrow mesenchymal stem cells were extracted from 21 healthy donors and 25 ankylosing spondylitis patients respectively, and passage 4 cells were used in subsequent experiments. A human monocytic cell line was induced to differentiate into macrophages. The phenotypic markers of bone marrow mesenchymal stem cells and macrophages were detected by flow cytometry. Expressions of tumor necrosis factor-α and tumor necrosis factor-α-stimulated gene 6（TSG-6） proteins in the supernatant of co-culture system were detected by ELISA. Quantitative real-time PCR was applied to detect the m RNA level of cytokines secreted by bone marrow mesenchymal stem cells and macrophages. RESULTS AND CONCLUSION： The typical mesenchymal stem cell surface markers were expressed in both bone marrow mesenchymal stem cells from healthy donors and patients with ankylosing spondylitis, and CD68 was detected positively in induced macrophages. The protein and m RNA levels of tumor necrosis factor-α secreted by macrophages co-cultured with bone marrow mesenchymal stem cells from patients with ankylosing spondylitis were obviously higher than those from healthy donors（P〈0.05）. TSG-6 secreted by bone marrow mesenchymal stem cells from patients with ankylosing spondylitis was lower than that by bone marrow mesenchymal stem cells from healthy donors in both RNA transcriptional and protein levels（P〈0.05）. Our study demonstrates that bone marrow mesenchymal stem cells from patients with ankylosing spondylitis shows abnormal immunoregulatory function on inhibiting
出处
《中国组织工程研究》
CAS
北大核心
2016年第1期13-19,共7页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金资助项目(81271951
81401850)~~
