摘要
目的:探讨芍药苷对膝骨性关节炎(KOA)模型大鼠炎症及核转录因子-κB(NF-κB)/核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)信号通路的影响。方法:取SD大鼠采用前交叉韧带离断法建立KOA模型,随机分为模型组、氨基葡萄糖组、芍药苷低剂量组、芍药苷高剂量组、芍药苷高剂量+佛波酯(PMA)(NF-κB激活剂)组,每组10只,另选10只大鼠仅切开膝关节囊作为对照组,以氨基葡萄糖、芍药苷和PMA分组处理后,采用机械刺激法与热辐射法检测大鼠关节疼痛症状;检测大鼠膝关节宽度、关节肿胀度和滑膜厚度;HE染色检测各组大鼠关节组织结构形态;ELISA检测大鼠关节液及血清炎症因子IL-1β、单核细胞趋化蛋白1(MCP-1)、IL-9水平;免疫印迹检测大鼠关节组织NF-κB/NLRP3信号通路相关蛋白表达。结果:与对照组相比,模型组大鼠关节组织结构发生明显损伤,机械痛阈值、热敏痛阈值显著降低(P<0.05),膝关节宽度、关节肿胀度和滑膜厚度、关节液及血清IL-1β、MCP-1、IL-9水平、关节组织p-NF-κB p65/NF-κB p65及NLRP3蛋白表达升高(P<0.05)。与模型组相比,氨基葡萄糖组、芍药苷低剂量组和芍药苷高剂量组大鼠关节组织损伤均减轻,机械痛阈值、热敏痛阈值升高(P<0.05),膝关节宽度、关节肿胀度和滑膜厚度值、关节液及血清IL-1β、MCP-1、IL-9水平、关节组织p-NF-κB p65/NF-κB p65及NLRP3蛋白表达降低(P<0.05);芍药苷高剂量组大鼠关节组织损伤相比芍药苷低剂量组进一步减轻,机械痛阈值、热敏痛阈值进一步升高(P<0.05),膝关节宽度、关节肿胀度和滑膜厚度值、关节液及血清IL-1β、MCP-1、IL-9水平、关节组织p-NF-κB p65/NF-κB p65及NLRP3蛋白表达进一步降低(P<0.05)。与芍药苷高剂量组相比,芍药苷高剂量+PMA组大鼠关节组织损伤加重,机械痛阈值、热敏痛阈值降低(P<0.05),膝关节宽度、关节肿胀度和滑膜厚度值、关节液及血清IL-1β、MCP-1、IL-9水平、关节组织p-NF-κB p65/NF-κB p65及NLRP3蛋白表达升高(P<0.05);氨基葡萄糖组大鼠各指标差异无统计学意义(P>0.05)。结论:芍药苷可通过下调NF-κB/NLRP3信号通路发挥抗炎作用,进而减轻KOA大鼠关节损伤,并缓解其关节疼痛症状。
Objective:To investigate impacts of paeoniflorin on inflammation and nuclear transcription factor-κB(NF-κB)/nucleotide-binding oligomeric domain-like receptor protein 3(NLRP3)signal pathway in knee osteoarthritis(KOA)model rats.Meth-ods:SD rats were randomly grouped into model group,glucosamine group,paeoniflorin low-dose group,paeoniflorin high-dose group,paeoniflorin high-dose+phorbol ester(PMA)(NF-κB activator)group,with 10 rats in each group,another 10 rats were regarded as control group,and only knee joint capsule was cut,after treatment with glucosamine,paeoniflorin and PMA,joint pain symptoms of rats were detected by mechanical stimulation method and thermal radiation method;knee joint width,joint swelling and synovial thickness were measured;HE staining was applied to detect joint tissue structure and morphology of rats in each group;levels of inflamma-tory factors IL-1β,monocyte chemoattractant protein 1(MCP-1)and IL-9 in joint fluid and serum of rats were detected by ELISA;and Western blot was applied to detect expressions of NF-κB/NLRP3 signal pathway related proteins in rat joint tissue.Results:Com-pared with control group,joint tissue structure of model group was significantly damaged,mechanical pain threshold and thermal sen-sitivity pain threshold were significantly lower(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were higher(P<0.05).Compared with model group,joint tissue damage of rats in glucosamine group,paeoniflorin low-dose group and paeoniflorin high-dose group was reduced,mechanical pain threshold and thermal sensitivity pain threshold were higher(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expressions of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were lower(P<0.05);joint tissue injury of rats in paeoniflorin high-dose group was further reduced compared with paeoniflorin low-dose group,mechanical pain threshold and thermal sensitivity pain threshold were further higher(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were further lower(P<0.05).Compared with paeoniflorin high-dose group,joint tissue damage of rats in paeoniflorin high-dose+PMA group was increased,mechanical pain threshold and thermal sensitivity pain threshold were lower(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were higher(P<0.05);there was no significant change in all indexes of rats in glucosamine group(P>0.05).Conclusion:Paeoniflorin can play an anti-inflammatory role by down-regulating the NF-κB/NLRP3 signal pathway,thus alleviating joint injury and joint pain symptoms of KOA rats.
作者
张仲博
郑福增
王上增
史栋梁
郭中华
王云飞
王振涛
ZHANG Zhongbo;ZHENG Fuzeng;WANG Shangzeng;SHI Dongliang;GUO Zhonghua;WANG Yunfei;WANG Zhentao(Department of OrthopedicsⅠ,College of Orthopedics and Traumatology,Henan University of Chinese Medicine,Henan Provincial Hospital of Traditional Chinese Medicine,the Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450053,China;Department of Orthopedics and Traumatology,Henan University of Chinese Medicine,Henan Provincial Hospital of Traditional Chinese Medicine,Department of Joints,the Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450053,China)
出处
《中国免疫学杂志》
北大核心
2025年第8期1826-1832,共7页
Chinese Journal of Immunology
基金
河南省中医药科学研究专项课题(2022ZY2017)
2021年河南省中医药拔尖人才培养项目资助[豫卫中医函(2021)15号]
河南省中医药科学研究专项课题(2023ZY2097)
2022年度河南省自然科学基金面上项目(232300420269)。
