摘要
目的:白藜芦醇被认为是治疗骨关节炎(osteoarthritis,OA)的潜在候选药物,但其保护机制仍有待阐明。本研究旨在研究白藜芦醇对高脂饮食(high-fat diet,HFD)诱导的肥胖相关OA中核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)/胱天蛋白酶-1(Caspase-1)/白介素-1β(interleukin-1β,IL-1β)轴的影响及磷脂酰肌醇3激酶(phosphoinositide 3-kinase,PI3K)/丝氨酸/苏氨酸蛋白激酶(serine/threonine-protein kinase,AKT)信号通路在其中的作用。方法:雄性C57BL/6小鼠用HFD或HFD+白藜芦醇喂养22周。收集膝关节,应用组织学、免疫组织化学和PCR检测软骨损伤程度和NLRP3,Caspase-1和IL-1β表达。在体外实验中,用IL-1β处理SW1353软骨肉瘤细胞以模拟OA状态,并使用PI3K抑制剂LY294002阻断PI3K/AKT信号通路,Western blot检测NLRP3,Caspase-1和IL-1β蛋白表达。结果:HFD喂养的小鼠软骨组织出现OA样损伤,而白藜芦醇处理能减轻这种损伤。HFD上调小鼠软骨组织NLRP3,Caspase-1和IL-1β的mRNA和蛋白表达,而白藜芦醇处理显著抑制这种上调作用。此外,LY294002能降低SW1353细胞受IL-1β刺激升高的NLRP3和Caspase-1的表达,白藜芦醇联合LY294002共同处理可进一步下调NLRP3和Caspase-1的表达。单独白藜芦醇处理或LY294002处理降低NLRP3和Caspase-1表达的作用没有差异。结论:肥胖相关OA小鼠关节软骨中NLRP3/Caspase-1/IL-1β轴被激活,白藜芦醇通过抑制其激活发挥保护作用,且其抑制机制与PI3K/AKT信号通路无关。
Objective:Resveratrol is considered a potential candidate for the treatment of osteoarthritis(OA),but the mechanism of protection remains to be elucidated.The aim of this study was to investigate the effect of resveratrol on the NLRP3/Caspase⁃1/IL⁃1βaxis in high⁃fat diet(HFD)induced obesity⁃related OA,and to explore whether the effect of resveratrol on this axis is related to the activation of PI3K/AKT signaling pathway.Methods:Male C57BL/6 mice were fed with a HFD or HFD plus resveratrol for 22 weeks.The knee joint samples were collected for histological,immunohistochemical,and PCR analyses.For in vitro experiments,SW1353 chondrosarcoma cells were treated with IL⁃1βto mimic OA state,and LY294002,an inhibitor of PI3K,was used to block the PI3K/AKT signaling pathway.Then western blot analysis was used to detect the levels of protein expression.Results:Mice fed with HFD presented OA⁃like lesions in cartilage tissues,while resveratrol treatment alleviated this lesion.HFD upregulated the mRNA and protein expressions of NLRP3,Caspase⁃1 and IL⁃1β,while additional resveratrol treatment downregulated them.Besides,the protein expression of NLRP3 and Caspase⁃1 decreased in the presence of LY294002 in IL⁃1β⁃stimulated SW1353 cells,and additional resveratrol further downregulated their expression.However,compared with IL⁃1β+RES group,LY294002 treatment had no significant effect on the expression of NLRP3 and Caspase⁃1.Conclusion:NLRP3/Caspase⁃1/IL⁃1βaxis was activated in articular cartilage of mice with obesity⁃associated OA,and resveratrol exerted protective effect by reversing its activation.The inhibitory mechanism is independent of the PI3K/AKT signaling pathway.
作者
王睿淇
何健宜
孙应许
杨迎春
顾海伦
刘莉
WANG Rui-qi;HE Jian-yi;SUN Ying-xu;YANG Ying-chun;GU Hai-lun;LIU Li(Key Laboratory of Environmental Stress and Chronic Disease Control&Prevention,China Medical University,Ministry of Education,Shenyang 110122,China;Department of Nutrition and Food Hygiene,School of Public Health,China Medical University,Shenyang 110122,China;Department of Orthopedics,Shengjing Hospital,China Medical University,Shenyang 110004,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2024年第17期1787-1795,共9页
Chinese Journal of New Drugs
基金
辽宁省教育厅科学研究经费面上项目(LJKZ0747)
辽宁省自然科学基金计划资助项目(2022-MS-093)。
