Zhuangguguaniie Formulation Protects Articular Cartilage From Degeneration In Joint Instability-induced Murine Knee Osteoarthritis

Backgroumd:Osteoarthritis(OA)is a slowly progressing,extremely prevalent disease—more than one-third of the world's adult population exhibits radiologic signs of OA.The pathological process of OA is characterized by progressive cartilage degeneration and subchondral bone sclerosis.These changes are due to a combination of mechanical and biological factors,which disrupt the normal balance between synthesis and degradation of chondrocytes,extracellular matrix components,and subchondral bone.At present,medical treatment of knee OA aims to relieve pain,reduce inflammation,delay cartilage degeneration,and improve function.However there is an extreme dearth of both clinical and fundamental research into CAM treatments for OA.As a time-tested traditional Chinese medicine,ZG is already in widespread use among China's OA patients.Weng Xisheng et al.have reported some degree of clinical success using ZG alone or in combination with celecoxib to treat OA,lumbar strains,and other conditions.Their results indicate that ZG has therapeutic potential for treating OA,but the authors did not explore its underlying mechanisms.Using the aforementioned research as a starting point,we conduct additional testing to determine the effectiveness of this traditional Chinese medicine,ZG,as a preventative treatment for knee OA.Methods:We conducted in vivo analysis on male ten-week-old C57BL/6 mice,using surgical destabilization of the medial meniscus(DMM)to model OA.Treatment groups were given either a solution containing ZG or a normal saline(NS)solution administered orally by gavage.At 4,6,and 8 weeks(post-surgery),surgically-manipulated knee joints were harvested and assessed with safranin-O staining,and blood serum was collected from the mice and tested for OA biomarkers.In vitro analysis was performed to evaluate cell toxicity of ZG and its effects on expression of OA-related genes.Finally,we used LC-MS to analyze the composition of the ZG solution.Results:We examined the effect of ZG on articular cartilage in the usage of both short-time and long-time.We found that the histological score of both femoral and tibia cartilage in ZG-treatment group is lower than that in the control one.At the same time,we detected the serum factors related to OA and found that the concentration of inflammatory cytokines,TNF-?,can be significantly reduced,as well as the biological marker of osteoclasts,CTX-1,after taking the ZG compound.It is suggested that the compound reagent can reduce the inflammatory reaction of the body and inhibit the activity of the osteoclasts,which may have the potential to treat osteoporosis.After that,we tested the effect of ZG solution on chondrocytes and found that it can significantly promote the expression of Collagen ? mRNA,and also inhibit the expression of MMP-3,13 and Adamts5.It suggests that the role of ZG could promote cartilage repair and inhibit the expression of extracellular matrix proteases.Finally,we found that ZG solution can inhibit the apoptosis of chondrocytes,and significantly promote the phosphorylation of AKT which inhibits the activation of Caspase-3.Moreover,under the action of AKT inhibitors,it can reverse the protection of chondrocytes by ZG solution.The effect suggests that Zhuanggu Joint Capsule can regulate the apoptosis of chondrocytes through the AKT-Caspase3 pathway and play a protective role in articular cartilage.Conclusion:Through the research of this topic,we used rigorous research methods to verify the protection of articular cartilage by Zhuanggu Joint Capsule,a commonly used compound for clinical treatment of osteoarthritis.And we clarify the possible inhibition of articular cartilage cells through the AKT-Caspase-3 pathway.At the same time,we also sought to search for new small molecules that protect articular cartilage in this research project,providing a new and effective potential treatment for patients with osteoarthritis.