TSP1 is the essential domain of SEMA5A involved in pannus formation in rheumatoid arthritis
TSP1 is the essential domain of SEMA5A involved in pannus formation in rheumatoid arthritis
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DOI：
10.1093/rheumatology/keab133
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发表时间：
2021-12-01
期刊：
影响因子：
5.5
通讯作者：
Yang, Xinyu
中科院分区：
文献类型：
--
作者：
Xiao, Chipeng;Lv, Chen;Yang, Xinyu
Objective.在这项研究中，我们探讨了信号蛋白5A（SEMA 5A）对RA发病机制的影响及其特异性TSP 1结构域对血管翳形成的影响。方法。采用实时荧光定量PCR（q-PCR）、免疫组化、Western blot和ELISA检测RA患者和健康对照组滑膜、滑膜成纤维细胞样细胞（FLS）和滑膜液（HC）中SEMA 5A的表达。进行SEMA 5A-mAb干预以评估CIA模型中关节的严重程度。对转染SEMA 5A的毛细胞FLS进行转录组测序和生物信息学分析，筛选SEMA 5A过表达后差异表达的基因。采用MTT法、鸡胚尿囊膜实验和试管形成实验研究SEMA 5A对RA-FLS细胞增殖和血管生成的影响。此外，拯救实验验证了SEMA 5A的TSP 1结构域在Sema 5a（-/-）CIA小鼠RA进展中的功能。SEMA 5A在RA中的表达明显高于HC。同时，SEMA 5A-mAb显著减弱CIA模型中的关节损伤和炎症反应。此外，转录组测序和血管生成相关实验验证了SEMA 5A促进FLS增殖和血管生成的能力。此外，TSP 1被证明是SEMA 5A在体外诱导血管生成的一个重要结构域。此外，在用Sema 5a（-/-）小鼠构建的CIA模型中，补救TSP 1缺失的SEMA 5A未能降低关节炎的严重程度。总之，SEMA 5A的上调首先在RA患者的病理学病变中得到证实。此外，在CIA模型中，用SEMA 5A-mAbs治疗减弱了RA的进展。此外，TSP 1被认为是SEMA 5A促进RA血管翳形成的关键结构域。[图形]。
Objective. In this study, we explored the effect of semaphorin5A (SEMA5A) on RA pathogenesis and its specific TSP1 domain on pannus formation.Methods. The expression of SEMA5A was detected in the synovium, the fibroblast-like synoviocytes (FLSs) and the SF of RA patients and healthy controls (HCs) by real-time quantitative PCR (q-PCR), immunohistochemistry staining, western blot and ELISA. SEMA5A-mAb intervention was performed to appraise the severity of joints in the CIA model. Transcriptome sequencing and bioinformatics analysis in SEMA5A-transfected FLSs from HCs were performed to screen differentially expressed genes after SEMA5A overexpression. An MTT assay in RA-FLSs, a chicken embryo allantoic membrane experiment and a tube formation experiment were used to clarify the influence of SEMA5A on cell proliferation and angiogenesis. Furthermore, a rescue experiment verified the function of the TSP1 domain of SEMA5A in the progress of RA with Sema5a(-/-) CIA mice.Results. The expression of SEMA5A increased in RA compared with that in HCs. Simultaneously, SEMA5A-mAbs significantly attenuated joint injury and the inflammatory response in CIA models. In addition, transcriptome sequencing and angiogenesis-related experiments verified the ability of SEMA5A to promote FLS proliferation and angiogenesis. Moreover, TSP1 was proved to be an essential domain in SEMA5A-induced angiogenesis in vitro. Additionally, rescue of TSP1-deleted SEMA5A failed to reduce the severity of arthritis in a CIA model constructed with Sema5a(-/-) mice.Conclusion. In summary, upregulation of SEMA5A was first confirmed in pathological lesions of RA patients. Furthermore, treatment with SEMA5A-mAbs attenuated the progress of RA in the CIA model. Moreover, TSP1 was indicated as the key domain of SEMA5A in the promotion of pannus formation in RA.[GRAPHICS].