Jiang Nan, Liu Hong-Xiao, Liang Hui-Ying, Feng Xing-Hua, Liu Ben-Yong, Zhou Ying-Yan
Department of Nephrology and Rheumatology, Second Affiliated Hospital of Tianjin University of TCM, Tianjin, China.
Department of Rheumatology, Guang'anmen Hospital, China Academy of Chinese Medical Sciences, Beijing, China.
Ann Transl Med. 2021 Feb;9(4):331. doi: 10.21037/atm-20-7817.
Autoimmune disease are fairly common and one that has an excessive degree of disability is Ankylosing spondylitis (AS). As the main cells in connective tissues, fibroblasts may play important roles in AS ossification. The conducted research aims to establish the osteogenic disparity characteristics of fibroblasts cultured , obtained via AS patients hip joint capsule, as well as investigating the pathological osteogenic molecular workings of AS.
AS patients hip joint capsules were acquired and fracture patients as the control with the finite fibroblast line were established by using tissue culture method. AS fibroblast proliferation, cycle and apoptosis, expression of osteogenic marker genes, osteogenic phenotypes, and the activation degree of the bone morphogenetic protein (BMP)/Smads signalling pathway were detected by flow cytometry, western blotting and real-time fluorescent quantitative polymerase chain reaction.
Proliferative activity in AS fibroblasts were abnormally high, and the apoptotic rate decreased. Compared with normal fibroblasts, the mRNA expression of osteogenic marker genes, expression of osteogenic phenotypes, protein expression of core-binding factor a1 (Cbfa1), Smad1, Smad4, Smad5, phosphorylated (p) Smad1, and pSmad5 in AS fibroblasts were higher; however, the expression of Smad6 was lower. Moreover, recombinant human bone morphogenetic protein-2(rhBMP-2) stimulated Cbfa1 expression by normal and AS fibroblasts through the BMP/Smads signalling pathway.
The fibroblasts of hip joint capsules in patients with AS cultured have biologic characteristics of osteogenic differentiation and may be important target cells of AS ossification. The Activated BMP/Smads signalling pathway could potentially be a mechanism relating to fibroblasts differentiating into osteoblasts and an ossification mechanism for AS.
自身免疫性疾病相当常见,其中强直性脊柱炎(AS)是一种致残程度较高的疾病。成纤维细胞作为结缔组织中的主要细胞,可能在AS骨化过程中发挥重要作用。本研究旨在确定通过AS患者髋关节囊培养获得的成纤维细胞的成骨差异特征,并研究AS的病理性成骨分子机制。
获取AS患者的髋关节囊,并以骨折患者为对照,采用组织培养方法建立有限的成纤维细胞系。通过流式细胞术、蛋白质印迹法和实时荧光定量聚合酶链反应检测AS成纤维细胞的增殖、周期和凋亡、成骨标记基因的表达、成骨表型以及骨形态发生蛋白(BMP)/Smads信号通路的激活程度。
AS成纤维细胞的增殖活性异常高,凋亡率降低。与正常成纤维细胞相比,AS成纤维细胞中成骨标记基因的mRNA表达、成骨表型的表达、核心结合因子a1(Cbfa1)、Smad1、Smad4、Smad5、磷酸化(p)Smad1和pSmad5的蛋白表达较高;然而,Smad6的表达较低。此外,重组人骨形态发生蛋白-2(rhBMP-2)通过BMP/Smads信号通路刺激正常和AS成纤维细胞中Cbfa1的表达。
培养的AS患者髋关节囊成纤维细胞具有成骨分化的生物学特性,可能是AS骨化的重要靶细胞。激活的BMP/Smads信号通路可能是成纤维细胞分化为成骨细胞的相关机制以及AS的骨化机制。