Wei Yibao, Ma Zhenyuan, Li Zhenhui, Kang Junfeng, Liao Taiyang, Jie Lishi, Liu Deren, Shi Lei, Wang Peimin, Mao Jun, Wu Peng
Department of Orthopaedics and Traumatology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China; Jiangsu Province Hospital of Chinese Medicine, Nanjing 210029, China; Key Laboratory for Metabolic Diseases in Chinese Medicine, First College of Clinical Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China.
Department of Orthopaedics and Traumatology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China; Jiangsu Province Hospital of Chinese Medicine, Nanjing 210029, China.
Int Immunopharmacol. 2025 Feb 6;147:113973. doi: 10.1016/j.intimp.2024.113973. Epub 2025 Jan 6.
Knee osteoarthritis (KOA) is a degenerative joint disease characterized by synovial inflammation and fibrosis. Gentiopicroside (GPS), one of the main active ingredients of Gentiana macrophylla, is widely used in anti-inflammatory and anti-fibrotic therapies. However, the exact mechanism by which GPS treats synovial inflammation and fibrosis in KOA remains unclear.
Fibroblast-like synoviocytes (FLSs) were stimulated with lipopolysaccharide (LPS) to induce inflammation and fibrosis, and CCK-8 was performed to determine the viability of GPS-treated FLSs, using immunofluorescence to examine the expression of P-PI3K and P-AKT, confocal microscopy was used to identify intracellular HMGB1 translocation. The KOA rat model was established by anterior cruciate ligament transection (ACLT) and subsequently subjected to GPS intervention. Inflammatory cytokines (TNF-α, IL-1β, and IL-6), fibrosis-related indicators (TGF-β, collagen I, TIMP1, and α-SMA), and HMGB1/PI3K/AKT signaling axis-related proteins and gene expression of fibroblast-like synoviocytes and synovial tissues were detected by Western blotting and real-time PCR. The histopathology of the synovium of the rats was assessed using Hematoxylin-eosin (HE), Sirius Red, and Masson staining. Immunohistochemistry was performed to detect the expression of HMGB1, P-PI3K, and P-AKT.
The present study revealed that GPS intervention significantly ameliorated inflammation and fibrosis in LPS-stimulated FLSs and KOA rat synovium. Immunofluorescence demonstrated that GPS inhibited the release of HMGB1 from the nucleus. Furthermore, GPS intervention down-regulates the levels of proteins and gene associated with the HMGB1/PI3K/AKT signaling pathway.
GPS ameliorated synovial inflammation and fibrosis in KOA rats, which may involve HMGB1-mediated activation of the PI3K/AKT signaling axis.
膝关节骨关节炎（KOA）是一种以滑膜炎症和纤维化为特征的退行性关节疾病。龙胆苦苷（GPS）是秦艽的主要活性成分之一，广泛用于抗炎和抗纤维化治疗。然而，GPS治疗KOA滑膜炎症和纤维化的确切机制尚不清楚。
用脂多糖（LPS）刺激成纤维样滑膜细胞（FLSs）以诱导炎症和纤维化，采用CCK-8法检测GPS处理的FLSs的活力，用免疫荧光法检测P-PI3K和P-AKT的表达，用共聚焦显微镜鉴定细胞内HMGB1易位。通过前交叉韧带横断术（ACLT）建立KOA大鼠模型，随后进行GPS干预。通过蛋白质免疫印迹法和实时荧光定量PCR检测炎性细胞因子（TNF-α、IL-1β和IL-6）、纤维化相关指标（TGF-β、I型胶原、TIMP1和α-SMA）以及成纤维样滑膜细胞和滑膜组织中HMGB1/PI3K/AKT信号轴相关蛋白和基因的表达。采用苏木精-伊红（HE）、天狼星红和Masson染色评估大鼠滑膜的组织病理学。进行免疫组织化学检测HMGB1、P-PI3K和P-AKT的表达。
本研究表明，GPS干预显著改善了LPS刺激的FLSs和KOA大鼠滑膜中的炎症和纤维化。免疫荧光显示，GPS抑制了HMGB1从细胞核中的释放。此外，GPS干预下调了与HMGB1/PI3K/AKT信号通路相关的蛋白质和基因水平。
GPS改善了KOA大鼠的滑膜炎症和纤维化，这可能涉及HMGB1介导的PI3K/AKT信号轴激活。
