Effect and mechanism of dichloromethane extract of fresh Sambucus adnata Wall.in rat osteoarthritis models

Lu Yuchun; Zhu Zimo; Li Chaomeng; Liu Ju; Jiang Zixian; Li Xiufang; Wang Tao; Wang Wenjing

doi:10.12307/2026.712

Abstract：BACKGROUND:The previous study conducted by the research group demonstrated that the methanol and dichloromethane extract of fresh Sambucus adnata Wall.(fSAW-DCE)exhibits anti-inflammatory,anti-oxidation,anti-angiogenesis properties,and shows significant anti-osteoarthritis activity. OBJECTIVE:To investigate the protective effects and underlying mechanisms of fSAW-DCE on osteoarthritis. METHODS:(1)The rat osteoarthritis model was replicated by anterior cruciate ligament transection.Following the operation,fSAW-DCE was administered intragastrically for a duration of 6 weeks.The concentration of tumor necrosis factor-α in the joint lavage fluid was evaluated.Hematoxylin-eosin staining was employed to observe the pathological changes of the articular cartilage,and the degree of mineralization of the cartilage was quantified.Micro-CT was used for imaging examination of osteophyte proliferation and changes in bone mass of the subchondral bone.(2)Insulin,transferrin,sodium selenite,and vitamin C were used to differentiate the mouse embryonic tumor cell line ATDC5 into chondrocytes.The chondrocyte inflammation model was replicated by interleukin-1β stimulation.Following this,the cells were treated with fSAW-DCE.The content of tumor necrosis-α in the cell supernatant was detected by ELISA kit,and the expression of Aggrecan and type II collagen mRNA was detected by RT-qPCR.(3)Transcriptome sequencing method was used to detect the differential genes after fSAW-DCE intervention in the ATDC5 cell inflammatory model,and to explore its mechanism. RESULTS AND CONCLUSION:(1)fSAW-DCE demonstrated the capacity to suppress joint inflammation,decrease the level of tumor necrosis factor-α in the joint fluid(P<0.05),retard mineralization of the articular cartilage(P<0.05),mitigate cartilage injury,and maintain subchondral bone volume(P<0.05).(2)fSAW-DCE could alleviate inflammation in ATDC5 chondrocytes induced by interleukin-1β(P<0.05)and preserve the mRNA expression of type II collagen in the extracellular matrix of chondrocytes(P<0.05).(3)Transcriptome sequencing and subsequent verification indicated that the anti-osteoarthritis effects of fSAW-DCE may be associated with inhibiting the tumor necrosis factor/nuclear factor-κB signaling pathway.