Anti-inflammatory and Anti-arthritic Effects of Standardized Hot Water Extract from Hydrangea serrata (Thunb.) Ser. via Modulation of NF-κB and MAPK Pathways in vitro and in vivo

다국어 초록 (Multilingual Abstract)

Osteoarthritis (OA) is a debilitating condition characterized by joint cartilage degeneration and inflammation. Although the anti-inflammatory properties of Hydrangea serrata (Thunb.) Ser. (H. serrata) have been known, its full potential as a treatment for OA, particularly using a standardized hot water extract of H. serrata (WHS), has not been thoroughly explored. This study was conducted to evaluate the anti-inflammatory and anti-arthritic properties of WHS in both cell-based models and animal models, focusing on its impact on the nuclear factor kappa-light-chain- enhancer of activated B cells (NF-κB) and mitogen-activated protein kinase (MAPK) pathways. In vitro, WHS reduced key inflammatory mediators (PGE2 and COX-2) and the mRNA expression of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in RAW 264.7 murine macrophages and SW1353 human chondrocytes. Furthermore, WHS effectively inhibited RANKL-induced osteoclastogenesis in RAW 264.7 macrophages, suggesting its potential in preventing bone degradation in OA. In vivo, WHS reduced paw swelling and inflammatory pain in carrageenan-induced paw edema and adjuvant-induced arthritis (AIA) rat models. These effects were associated with decreased phosphorylation of IκB kinase alpha/beta (IKKα/β) and MAPKs, as well as reduced expression of metalloproteinase-13 (MMP-13). These findings suggest that WHS is a promising therapeutic agent for OA and its associated inflammatory conditions.

목차 (Table of Contents)

• Abstract v
• 1. Introduction 1
• 2. Material and Methods 4
• 2.1. Materials 4
• 2.2. Cell culture and cytotoxicity assay 5
• 2.3. NO assay 6
• 2.4. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis 6
• 2.5. Western blot analysis 8
• 2.6. Osteoclast differentiation and TRAP staining 9
• 2.7. Animals 9
• 2.8. Establishing carrageenan-induced paw edema model 10
• 2.9. Establishing adjuvant-induced arthritis model 11
• 2.10. Histological analysis 12
• 2.11. Micro-CT analysis 12
• 2.12. Statistical analysis 12
• 3. Results 13
• 3.1. WHS exhibits anti-inflammatory effects in LPS-induced RAW 264. 7 macrophages 13
• 3.2. WHS inhibits osteoclastogenesis in RANKL-induced RAW 264. 7 macrophages 16
• 3.3. WHS exhibits anti-arthritic effects in IL-1β-induced SW 1353 chondrocytes 18
• 3.4. WHS inhibits NF-κB and MAPK pathways in IL-1β-induced SW 1353 chondrocytes 20
• 3.5. WHS reduces acute inflammation in carrageenan-induced paw edema rats 22
• 3.6. WHS demonstrates anti-arthritic efficacy in an adjuvant-induced arthritis rats 24
• 3.7. WHS suppresses inflammatory mediators in the synovial tissues of adjuvant-induced arthritis rats 26
• 3.8. WHS inhibited cartilage degradation in adjuvant-induced arthritis rats · 27
• 3.9. Micro-CT analysis in adjuvant-induced arthritis rats 29
• 4. Discussion 30
• 5. Conclusion 36
• 6. References 37