Department of Orthopedics, the First Affiliated Hospital of Anhui Medical University, Hefei, 230000, China
Jt Dis Relat Surg. 2024 Jul 8;35(3):513-520. doi: 10.52312/jdrs.2024.1247.
This study aims to explore the mechanisms of dual regulation of osteoarthritis (OA) progression by the involvement of estrogen receptor (ER) in autophagy and inflammation.
Bioinformatics methods were used to explore the relationship among associated genes. Western blot assays were used to detect related protein expression of OA in C28I2 and induced OA cellular model. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis were used to detect OA related gene expression in C28I2 and induced OA cellular model. Co-immunoprecipitation (CO-IP) analysis were used to verify the direct interaction between ER and NOD-like receptor thermal protein domain associated protein 3 (NLRP3).
The C28I2 cellular model of OA was induced by interleukin-1β (IL-1β). The small interfering ribonucleic acid (SiRNA)-mediated knockdown of autophagy-related 16 like 1 (ATG16L1) in C28I2 decreased the expression of MAP1LC3B (LC3B) and NLRP3. Besides, ER-beta (ERβ) agonist changed the gene expression of NLRP3 and ATG16L1. Moreover, CO-IP analysis indicated the direct interaction between ER and NLRP3.
Our study results revealed that ATG16L1, NLRP3, and IL-1β interacted closely and ERβ was involved in OA process by affecting autophagy and inflammatory activation.
本研究旨在探讨雌激素受体（ER）在自噬和炎症参与下对骨关节炎（OA）进展的双重调节机制。
采用生物信息学方法探讨相关基因之间的关系。采用 Western blot 检测 C28I2 和诱导 OA 细胞模型中 OA 相关蛋白的表达。采用实时定量聚合酶链反应（RT-qPCR）分析 C28I2 和诱导 OA 细胞模型中 OA 相关基因的表达。采用免疫共沉淀（CO-IP）分析验证 ER 与 NOD 样受体热蛋白结构域相关蛋白 3（NLRP3）之间的直接相互作用。
采用白细胞介素-1β（IL-1β）诱导 C28I2 建立 OA 细胞模型。C28I2 细胞中转染自噬相关基因 16 样 1（ATG16L1）的小干扰 RNA（siRNA）可降低 MAP1LC3B（LC3B）和 NLRP3 的表达。此外，ER-β（ERβ）激动剂改变了 NLRP3 和 ATG16L1 的基因表达。此外，CO-IP 分析表明 ER 与 NLRP3 之间存在直接相互作用。
本研究结果表明，ATG16L1、NLRP3 和 IL-1β 相互作用密切，ERβ 通过影响自噬和炎症激活参与 OA 进程。
