南蛇藤素阻断"TLR4-HIF-1α"对话抗RA滑膜细胞侵袭的机制

Fibroblast-like synoviocyte (FLS) invasion is the primary cause of joint destruction in rheumatoid arthritis (RA). TLR4 and HIF-1α are closely related to cell invasion. Cross-talk may exist among them with cell difference. Our early-stage research demonstrates that celastrol can inhibit signal pathway activation of TLR4/NF-κB and can restrain the FLS invasion stimulated by LPS. Preliminary experiment also found that celastrol can inhibit FLS invasion and TLR4-HIF-1α expression induced by hypoxia and LPS . However, the cross-talk between TLR4 and HIF-1α in celastrol inhibiting FLS invasion or the role in it has not been reported yet. Therefore, it can be hypothesized that celastrol inhibit FLS invasion by blocking the cross-talk between TLR4 and HIF-1α. This paper intends to utilize the technology as RNA interference, dual Luciferase reporter genes, Electrophoretic Mobility Shift Assay (EMSA) and Chromatin immunoprecipitation(ChIP) , takes SCID-HuRAg mice and FLS as the object of study, systemically explores whether celastrol plays the role of resisting FLS invasion by blocking the cross-talk between TLR4 and HIF-1α, and reveals the mechanism of Celastrol resisting FLS invasion from the whole to cellular level, so as to provide experiment basis for celastrol treating RA and usher in a new path of exploring RA new-type treatment strategy.

滑膜成纤维细胞（FLS）侵袭是类风湿关节炎（RA）关节破坏的主因。TLR4、HIF-1α与细胞侵袭密切相关，他们之间可能存在对话，且具有细胞差异性。我们前期研究证实南蛇藤素可阻断TLR4/NF-κB信号通路活化，抑制LPS介导的FLS侵袭。预实验也发现，南蛇藤素可抑制低氧和LPS共同介导的FLS侵袭以及TLR4-HIF-1α表达，然TLR4与HIF-1α对话在南蛇藤素抑制FLS侵袭中的作用机制尚未明确。由此，我们假设：南蛇藤素阻断TLR4和HIF-1α对话是其抗FLS侵袭的重要机制。拟利用RNA干扰、双荧光素酶报告基因、EMSA及ChIP等技术方法，分别以SCID-HuRAg小鼠及FLS为研究对象，系统探讨南蛇藤素是否通过阻断TLR4和HIF-1α对话而发挥抗FLS侵袭作用，从整体-细胞水平揭示南蛇藤素抗FLS侵袭的机制，为南蛇藤素治疗RA提供实验依据，也为探索RA新型治疗策略开辟思路。

滑膜成纤维细胞（FLS）侵袭是类风湿关节炎（RA）关节破坏的主因。TLR4、HIF-1α与细胞侵袭密切相关，他们之间可能存在对话，且具有细胞差异性。我们前期研究证实南蛇藤素可阻断TLR4/NF-κB信号通路活化，抑制LPS介导的FLS侵袭。预实验也发现，南蛇藤素可抑制低氧和LPS共同介导的FLS侵袭以及TLR4-HIF-1α表达，然TLR4与HIF-1α对话在南蛇藤素抑制FLS侵袭中的作用机制尚未明确。由此，我们假设：南蛇藤素阻断TLR4和HIF-1α对话是其抗FLS侵袭的重要机制。拟利用RNA干扰、双荧光素酶报告基因、EMSA及ChIP等技术方法，分别以SCID-HuRAg小鼠及FLS为研究对象，系统探讨南蛇藤素抗FLS侵袭作用及机制，结果发现，南蛇藤素可通过阻断TLR4和HIF-1α对话发挥其抗RA-FLS侵袭及抗RA骨侵蚀效应，从整体-细胞水平揭示南蛇藤素抗FLS侵袭的机制，为南蛇藤素治疗RA提供实验依据，也为探索RA新型治疗策略开辟思路。