Mei Jiajun, Ke Hongkui, Zhu Junsong
Department of Pain Medicine, Wuhan University of Science and Technology Affiliated Puren Hospital, Wuhan, China.
Cell Biochem Biophys. 2025 Jan 7. doi: 10.1007/s12013-024-01635-9.
Icariside II exerts protective effects against various diseases; however, its specific effects on osteoarthritis (OA) remain unclear. Therefore, in this study, we aimed to investigate the effects of icariside II in an in vitro model of OA and analyze its action mechanisms. We established an in vitro OA model by treating a human chondrocyte cell line (CHON-001) with interleukin (IL)-1β, followed by treatment with different concentrations of icariside II. Cell viability was measured using the methyl thiazolyl tetrazolium assay, and the level of lactate dehydrogenase (LDH) released from cells was determined using the appropriate kit. Tumor necrosis factor (TNF)-α, IL-6, and IL-8 levels were determined via enzyme-linked immunosorbent assay. Flow cytometry was used to assess apoptosis. Apoptosisrelated protein expression levels and TNFAIP3-interacting protein 2 (TNIP2)/nuclear factor (NF)-κB signaling pathway were analyzed via reverse transcription-quantitative polymerase chain reaction and western blotting. Furthermore, TNIP2-small interfering RNA (siRNA) was used to determine whether the TNIP2/NF-κB pathway influences the effects of icariside II on OA. Results indicated that Icariside II did not exert any significant toxic effects on CHON-001 cells. It inhibited IL-1β-induced apoptosis and increase in LDH levels and enhanced the inflammatory response. Additionally, icariside II reversed the IL-1β-induced decrease in TNIP2 levels and increase in NF-κB phosphorylation. TNIP2-siRNA revealed that the TNIP2/NF-κB signaling pathway influenced the alleviating effects of icariside II on OA. In conclusion, our results revealed that icariside II attenuated IL-1β-induced inflammatory injury in chondrocytes by increasing TNIP2 expression and inhibiting NF-κB pathway activation, highlighting its therapeutic potential for OA.
淫羊藿次苷II对多种疾病具有保护作用；然而，其对骨关节炎（OA）的具体作用仍不清楚。因此，在本研究中，我们旨在研究淫羊藿次苷II在OA体外模型中的作用，并分析其作用机制。我们通过用白细胞介素（IL）-1β处理人软骨细胞系（CHON-001）建立了OA体外模型，随后用不同浓度的淫羊藿次苷II进行处理。使用甲基噻唑基四氮唑法测定细胞活力，并使用相应试剂盒测定细胞释放的乳酸脱氢酶（LDH）水平。通过酶联免疫吸附测定法测定肿瘤坏死因子（TNF）-α、IL-6和IL-8水平。使用流式细胞术评估细胞凋亡。通过逆转录-定量聚合酶链反应和蛋白质印迹分析凋亡相关蛋白表达水平以及TNFAIP3相互作用蛋白2（TNIP2）/核因子（NF）-κB信号通路。此外，使用TNIP2小干扰RNA（siRNA）来确定TNIP2/NF-κB通路是否影响淫羊藿次苷II对OA的作用。结果表明，淫羊藿次苷II对CHON-001细胞没有显著毒性作用。它抑制IL-1β诱导的细胞凋亡和LDH水平升高，并增强炎症反应。此外，淫羊藿次苷II逆转了IL-1β诱导的TNIP2水平降低和NF-κB磷酸化增加。TNIP2-siRNA显示TNIP2/NF-κB信号通路影响淫羊藿次苷II对OA的缓解作用。总之，我们的结果表明，淫羊藿次苷II通过增加TNIP2表达和抑制NF-κB通路激活减轻IL-1β诱导的软骨细胞炎症损伤，突出了其对OA的治疗潜力。
