Abstract：
Osteoarthritis (OA) is a heterogeneous disease that affects the entire joint. Its pathogenesis involves hypertrophy and hyperplasia of synovial cells and polarization infiltration of macrophages, in which macrophages, as a potential target, can delay the progression of the disease by improving the immune microenvironment in OA. To investigate the role and regulatory mechanism of Carveol in cartilage and synovial macrophage reprogramming and crosstalk during the development of OA. RAW264.7 mouse macrophage cell line was mainly used to stimulate macrophages to polarization towards M1 and M2 by LPS, IL4+IL13, respectively. Different concentrations of Carveol were given to intervene, and macrophage culture medium was collected to intervene mouse C57BL6J chondrocytes. ROS assay kit, western blotting, cellular immunofluorescence, scanning microscope and section histology were used to evaluate the effect of Carveol on anti-M1-polarization, M2-polarization promotion and cartilage protection. The mouse destabilization of medial meniscus (DMM) model was observed by micro-CT scan and histology. We found that CA could inhibit the increase of macrophage inflammation level under the intervention of LPS and promote the production of M2 anti-inflammatory substances under the intervention of IL-4+IL13. In addition, Carveol activated NRF2/HO-1/NQO1 pathway and enhanced ROS clearance in chondrocytes under the intervention of macrophage culture medium. The phosphorylation of I-κBα is inhibited, which further reduces the phosphorylation of P65 downstream of nuclear factor-κB (NF-κB) signaling pathway. In addition, Carveol inhibits mitogen activated protein kinase (MAPK) signaling molecules P-JNK, P-ERK and P-P38, and inhibits the production of inflammatory mediators. In vivo, Carveol can reduce osteophytes and bone spurs induced by DMM, reduce hypertrophy of synovial cells, reduce infiltration of macrophages, inhibit subchondral bone destruction, and reduce articular cartilage erosion. Our study suggests that synovial macrophages are potential targets for OA treatment, and Carveol is an effective candidate for OA treatment.
摘要：
骨关节炎（OA）是一种影响整个关节的异质性疾病。其发病机制涉及滑膜细胞肥大增生和巨噬细胞极化浸润,其中巨噬细胞，作为一个潜在的目标,可以通过改善OA的免疫微环境来延缓疾病的进展。探讨卡维醇在OA发生发展过程中软骨和滑膜巨噬细胞重编程和串扰中的作用及调控机制。RAW264.7小鼠巨噬细胞系主要用于通过LPS刺激巨噬细胞向M1和M2极化,分别为IL4+IL13。给予不同浓度的Carveol进行干预，收集巨噬细胞培养液干预小鼠C57BL6J软骨细胞。ROS测定试剂盒,西方印迹,细胞免疫荧光，扫描显微镜和切片组织学用于评估Carveol对抗M1极化的影响，M2极化促进和软骨保护。通过显微CT扫描和组织学观察小鼠内侧半月板（DMM）模型的失稳。我们发现CA能在LPS的干预下抑制巨噬细胞炎症水平的升高,在IL-4+IL13的干预下促进M2类抗炎物质的产生。此外,Carveol激活NRF2/HO-1/NQO1通路，并在巨噬细胞培养基的干预下增强软骨细胞中的ROS清除。I-κBα的磷酸化被抑制,这进一步降低了核因子-κB（NF-κB）信号通路下游P65的磷酸化。此外,卡维醇抑制丝裂原活化蛋白激酶（MAPK）信号分子P-JNK，P-ERK和P-P38，并抑制炎症介质的产生。在体内,Carveol可以减少DMM引起的骨赘和骨刺，减少滑膜细胞肥大，减少巨噬细胞的浸润，抑制软骨下骨破坏，减少关节软骨侵蚀。我们的研究表明滑膜巨噬细胞是治疗OA的潜在靶点，Carveol是治疗OA的有效候选药物。
