Department of Sports Medical, Ganzhou People's Hospital/The Affiliated Ganzhou Hospital of Nanchang University, Ganzhou, china.
Cell Cycle. 2021 Jun;20(12):1107-1121. doi: 10.1080/15384101.2021.1916272. Epub 2021 Jun 7.
Studies have found that cell pyroptosis is involved in the occurrence and development of rheumatoid arthritis (RA). Hsa_circ_0044235 has been found to be significantly low-expressed in RA patients. The purpose of this research was to reveal the regulatory mechanism of hsa_circ_0044235 in the pyroptosis pathway of RA. Serum expressions of hsa_circ_0044235 and SIRT were detected by RT-qPCR, and the relationship of the two genes was analyzed by . Next, a collagen-induced arthritis (CIA) mouse model was constructed to examine the effect of hsa_circ_0044235 on knee joint injury. The number of apoptotic cells and the level of inflammatory cytokines in synovial tissue were detected by TUNEL and ELISA. Fibroblast-like synoviocytes (FLSs) were extracted as in vitro study subject. Functional assays including flow cytometry and immunofluorescence staining, molecular experiments including RT-qPCR, Western blot and dual luciferase assay, and bioinformatics analysis were performed to analyze the mechanism of hsa_circ_0044235 in pyroptosis in FLSs. Hsa_circ_0044235 and SIRT1 expressions were suppressed in RA patients and the two were positively correlated. Overexpressed hsa_circ_0044235 attenuated joint inflammation, cell apoptosis, and joint damage, reduced foot pad thickness, clinical case scores, inhibited the NLRP3-mediated pyroptosis pathway but promoted SIRT1 expression in CIA mice. Overexpressed hsa_circ_0044235 inhibited caspase-1 content and the NLRP3-mediated pyroptosis pathway. Moreover, hsa_circ_0044235 promoted SIRT1 expression by sponging miR-135b-5p in FLSs. Additionally, the effect of overexpressed hsa_circ_0044235 on FLSs was reversed by miR-135b-5p mimic and siSIRT1, while the effect of siSIRT1 was reversed by miR-135b-5p inhibitor. Hsa_circ_0044235 regulated NLRP3-mediated pyroptosis through miR-135b-5p-SIRT1 axis to regulate the development of RA.
研究发现细胞焦亡参与类风湿关节炎（RA）的发生发展。已经发现 hsa_circ_0044235 在 RA 患者中表达显著降低。本研究旨在揭示 hsa_circ_0044235 在 RA 焦亡通路中的调控机制。通过 RT-qPCR 检测 hsa_circ_0044235 和 SIRT 的血清表达，并通过 分析这两个基因之间的关系。接下来，构建胶原诱导性关节炎（CIA）小鼠模型，以检测 hsa_circ_0044235 对膝关节损伤的影响。通过 TUNEL 和 ELISA 检测滑膜组织中凋亡细胞的数量和炎症细胞因子的水平。提取成纤维样滑膜细胞（FLS）作为体外研究对象。进行流式细胞术和免疫荧光染色等功能测定、RT-qPCR、Western blot 和双荧光素酶测定等分子实验以及生物信息学分析，以分析 hsa_circ_0044235 在 FLS 中焦亡的机制。RA 患者中 hsa_circ_0044235 和 SIRT1 的表达受到抑制，且两者呈正相关。过表达 hsa_circ_0044235 可减轻关节炎症、细胞凋亡和关节损伤，降低足垫厚度、临床病例评分，抑制 CIA 小鼠 NLRP3 介导的焦亡通路，但促进 SIRT1 表达。过表达 hsa_circ_0044235 抑制 caspase-1 含量和 NLRP3 介导的焦亡通路。此外，hsa_circ_0044235 通过海绵吸附 miR-135b-5p 在 FLSs 中促进 SIRT1 表达。此外，过表达 hsa_circ_0044235 对 FLSs 的作用可以通过 miR-135b-5p 模拟物和 siSIRT1 逆转，而 siSIRT1 的作用可以通过 miR-135b-5p 抑制剂逆转。hsa_circ_0044235 通过 miR-135b-5p-SIRT1 轴调节 NLRP3 介导的焦亡，从而调节 RA 的发展。
