【摘要】 背景：课题组前期研究结果显示彝药斯赤列鲜品甲醇及二氯甲烷提取物具有抗炎、抗氧化、抗血管新生等作用，抗骨关节炎活性明显。目的：探究彝药斯赤列(鲜品)二氯甲烷提取物(fSAW-DCE)对骨关节炎的保护作用及机制。方法：(1)前交叉韧带切断法复制大鼠骨关节炎模型，术后连续灌胃斯赤列二氯甲烷提取物干预6周，ELISA检测关节腔液中肿瘤坏死因子α水平，苏木精-伊红染色观察关节软骨病理变化，计算关节软骨矿化程度，Micro CT影像学检查骨赘增生及软骨下骨骨量变化。(2)以小鼠胚胎瘤细胞ATDC5为研究对象，用胰岛素-转铁蛋白-亚硒酸钠和维生素C将其诱导分化为软骨细胞，用白细胞介素1β刺激复制软骨细胞炎症模型，然后给予斯赤列二氯甲烷提取物干预，ELISA试剂盒检测细胞上清液中肿瘤坏死因子α水平，RT-qPCR检测蛋白聚糖及Ⅱ型胶原mRNA表达。(3)转录组测序方法检测斯赤列二氯甲烷提取物干预软骨细胞炎症模型后的差异基因，探究作用机制。结果与结论：(1)斯赤列二氯甲烷提取物可抑制关节炎症，降低关节腔液中肿瘤坏死因子α水平(P<0.05)，延缓关节软骨矿质化(P<0.05)，减轻关节软骨损伤，维持软骨下骨骨量(P<0.05)；(2)斯赤列二氯甲烷提取物可减轻白细胞介素1β诱导的软骨细胞炎症(P<0.05)，维持软骨细胞外基质中Ⅱ型胶原mRNA表达水平(P<0.05)；(3)转录组学测序及验证结果显示斯赤列二氯甲烷提取物抗骨关节炎的作用与抑制肿瘤坏死因子/核因子κB信号通路的激活有关。更多还原

【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.更多还原