School and Hospital of Stomatology, China Medical University, Shenyang, Liaoning, China.
Department of Oral and Maxillofacial Surgery, Liaoning Provincial Key Laboratory of Oral Disease, Shenyang, Liaoning, China.
J Cell Mol Med. 2024 Apr;28(8):e18244. doi: 10.1111/jcmm.18244.
To explore the mechanism of tripartite motif 52 (TRIM52) in the progression of temporomandibular joint osteoarthritis (TMJOA). Gene and protein expression were tested by quantitative real-time polymerase chain reaction and western blot, respectively. The levels of pro-inflammatory cytokines and oxidative stress factors were evaluated using enzyme-linked immunosorbent assay and biochemical kit, respectively. Cell counting kit-8 and 5-ethynyl-2'-deoxyuridine assays were carried out to assess cell proliferation. Immunofluorescence was used to detect the expression of CD68 and Vimentin in primary synovial fibroblasts (SFs). Haematoxylin and eosin staining and Safranin O/Fast green were used to evaluate the pathological damage of synovial and cartilage tissue in rats. TRIM52 was upregulated in the synovial tissue and SFs in patients with TMJOA. Interleukin (IL)-1β treatment upregulated TRIM52 expression in TMJOA SFs and normal SF (NSF), promoting cell proliferation, inflammatory response and oxidative stress in NSF, SFs. Silence of TRIM52 relieved the cell proliferation, inflammatory response and oxidative stress induced by IL-1β in SFs, while overexpression of TRIM52 enhanced IL-1β induction. Meanwhile, IL-1β induction activated toll-like receptor 4 (TLR4)/nuclear factor (NF)-κB pathway, which was augmented by upregulation of TRIM52 in NSF, and was attenuated by TRIM52 knockdown in SFs. Besides, pyrrolidinedithiocarbamic acid ameliorated IL-1β-induced proliferation and inflammatory response by inhibiting TLR4/NF-κB signalling. Meanwhile, TRIM52 knockdown inhibited cell proliferation, oxidative stress and inflammatory response in IL-1β-induced SFs through downregulation of TLR4. TRIM52 promoted cell proliferation, inflammatory response, and oxidative stress in IL-1β-induced SFs. The above functions were mediated by the activation of TLR4/NF- κB signal pathway.
探讨三结构域蛋白 52（TRIM52）在颞下颌关节骨关节炎（TMJOA）进展中的作用机制。分别采用实时定量聚合酶链反应和蛋白质印迹法检测基因和蛋白质的表达。采用酶联免疫吸附试验和生化试剂盒分别评估促炎细胞因子和氧化应激因子的水平。通过细胞计数试剂盒-8 和 5-乙炔基-2'-脱氧尿苷检测来评估细胞增殖。免疫荧光法检测原代滑膜成纤维细胞（SFs）中 CD68 和波形蛋白的表达。苏木精和伊红染色和番红 O/快绿染色用于评估大鼠滑膜和软骨组织的病理损伤。TRIM52 在 TMJOA 患者的滑膜组织和 SFs 中上调。IL-1β处理上调 TMJOA SFs 和正常 SF（NSF）中的 TRIM52 表达，促进 NSF、SFs 中的细胞增殖、炎症反应和氧化应激。沉默 TRIM52 可缓解 SFs 中由 IL-1β引起的细胞增殖、炎症反应和氧化应激，而过表达 TRIM52 则增强了 IL-1β的诱导作用。同时，IL-1β诱导激活了 TLR4/核因子（NF）-κB 通路，TRIM52 在 NSF 中的上调增强了该通路，而在 SFs 中的下调则减弱了该通路。此外，吡咯烷二硫代氨基甲酸盐通过抑制 TLR4/NF-κB 信号通路改善了 IL-1β诱导的增殖和炎症反应。同时，TRIM52 敲低抑制了 IL-1β诱导的 SFs 中的细胞增殖、氧化应激和炎症反应，通过下调 TLR4。TRIM52 促进了 IL-1β诱导的 SFs 中的细胞增殖、炎症反应和氧化应激。上述功能是通过激活 TLR4/NF-κB 信号通路介导的。
