Department of Rheumatism and Immunology, Liaocheng People's Hospital, Liaocheng, 252000 Shandong, China.
Department of Orthopedics, Liaocheng Hospital of Traditional Chinese Medicine, Liaocheng, 252000 Shandong, China.
Mediators Inflamm. 2022 Nov 10;2022:4258742. doi: 10.1155/2022/4258742. eCollection 2022.
This study is aimed at investigating the role of -galactoside-2,3-sialyltransferase III (ST3GAL3) in fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA), as well as its potential mechanism of action. The Gene Expression Omnibus (GEO) database and gene set enrichment analysis (GSEA) were used to analyse the expression of and the enrichment signalling pathways associated with in RA. The effects of ST3GAL3 on tumour necrosis factor- (TNF-) and interleukin- (IL-) 1-treated MH7A cells were determined using methyl thiazolyl tetrazolium (MTT), transwell, and enzyme-linked immunosorbent assays (ELISA). The expression of proliferation-associated proteins and Toll-like receptor (TLR) pathway-enriched proteins was analysed using western blotting. As a main result, was screened as an overlapping upregulated gene from GSE101193 and GSE94519 datasets. ST3GAL3 expression in MH7A cells significantly increased with increasing treatment time with TNF- or IL-1. TLR9/myeloid differentiation primary response protein 88 (MyD88) is a downstream activation pathway of ST3GAL3. ST3GAL3 overexpression promoted MH7A cell proliferation and migration. Additionally, ST3GAL3 overexpression upregulated the expression of proliferation-associated proteins (cyclinD, cyclinE, and proliferating cell nuclear antigen) and TLR pathway enrichment factors (TLR9 and MyD88) and increased the production of matrix metallopeptidase (MMP) 1, MMP3, interleukin- (IL-) 6, and IL-8, whereas si-ST3GAL3 had the opposite effect. The addition of TLR9 agonists (CpG 2216 and CpG 2006) reversed the effects of si-ST3GAL3 on MH7A cell proliferation, migration, and inflammation. TLR9-specific siRNA reversed the effects of ST3GAL3 overexpression on MH7A cell proliferation, migration, and inflammation. In conclusion, ST3GAL3 is likely involved in RA pathogenesis by activating the TLR9/MyD88 pathway.
本研究旨在探讨β-半乳糖苷-2,3-唾液酸转移酶 III（ST3GAL3）在类风湿关节炎（RA）成纤维样滑膜细胞（FLS）中的作用及其潜在的作用机制。使用基因表达综合数据库（GEO）和基因集富集分析（GSEA）分析 RA 中与 相关的表达和富集信号通路。使用甲基噻唑基四唑（MTT）、Transwell 和酶联免疫吸附测定（ELISA）测定 ST3GAL3 对肿瘤坏死因子-（TNF-）和白细胞介素-（IL-）1 处理的 MH7A 细胞的影响。使用 Western blot 分析增殖相关蛋白和 Toll 样受体（TLR）通路富集蛋白的表达。作为主要结果，从 GSE101193 和 GSE94519 数据集筛选出重叠上调基因。随着 TNF-或 IL-1 处理时间的增加，MH7A 细胞中 ST3GAL3 的表达显著增加。TLR9/髓样分化初级反应蛋白 88（MyD88）是 ST3GAL3 的下游激活途径。ST3GAL3 过表达促进 MH7A 细胞增殖和迁移。此外，ST3GAL3 过表达上调增殖相关蛋白（cyclinD、cyclinE 和增殖细胞核抗原）和 TLR 通路富集因子（TLR9 和 MyD88）的表达，并增加基质金属蛋白酶（MMP）1、MMP3、白细胞介素-（IL-）6 和 IL-8 的产生，而 si-ST3GAL3 则有相反的作用。TLR9 激动剂（CpG 2216 和 CpG 2006）的添加逆转了 si-ST3GAL3 对 MH7A 细胞增殖、迁移和炎症的影响。TLR9 特异性 siRNA 逆转了 ST3GAL3 过表达对 MH7A 细胞增殖、迁移和炎症的影响。综上所述，ST3GAL3 通过激活 TLR9/MyD88 通路参与 RA 的发病机制。
