摘要
目的 探讨钩藤碱对IL-1β诱导的软骨细胞凋亡、氧化应激和炎症等损伤的影响及其分子机制。方法 将大鼠软骨细胞分为对照组、IL-1β(10 ng/mL)组、IL-1β+钩藤碱(5、25、50μmol/L)组;MTT实验检测钩藤碱对细胞活性的影响;Western blot方法检测Bcl2、Bax、MMP3、MMP13、CollagenⅡ、Nrf2和HO-1蛋白水平;ELISA方法检测炎性因子PGE2、COX-1、TNF-α和ROS水平;试剂盒检测MDA水平。结果 与对照组相比,IL-1β组细胞活性明显抑制(P<0.05);与IL-1β组相比,钩藤碱25μmol/L组和50μmol/L组细胞活性增强(P<0.05)。与对照组相比,IL-1β组Bcl2蛋白表达减少,Bax蛋白表达增加(P<0.05);与IL-1β组相比,钩藤碱25μmol/L组和50μmol/L组Bcl2蛋白表达增加,钩藤碱50μmol/L组Bax蛋白表达减少(P<0.05)。与对照组相比,IL-1β组MMP13和MMP3蛋白表达上调,CollagenⅡ蛋白表达下调(P<0.05);与IL-1β组相比,钩藤碱5、25和50μmol/L组MMP13和MMP3蛋白表达降低,钩藤碱25μmol/L组和50μmol/L组CollagenⅡ蛋白表达增加(P<0.05)。与对照组相比,IL-1β组PGE2、COX-1、TNF-α、ROS和MDA水平均升高(P<0.05);与IL-1β组相比,钩藤碱25μmol/L组和50μmol/L组PGE2、COX1、TNF-α、ROS和MDA水平均降低(P<0.05)。与对照组相比,IL-1β组Nrf2和HO-1蛋白表达减少(P<0.05);与IL-1β组相比,钩藤碱25μmol/L组和50μmol/L组Nrf2和HO-1蛋白表达增加(P<0.05)。结论 钩藤碱通过激活Nrf2/HO-1信号通路抑制IL-1β诱导的软骨细胞凋亡、氧化应激和炎症等损伤。
Objective To explore the effects of rhynchophylline on the oxidative stress and inflammatory injuries in IL-1β-induced chondrocytes.Methods The rat chondrocytes were divided into control group,IL-1β(10 ng/mL)group,and IL-1β+rhynchophylline(5,25,50μmol/L)groups.The effect of rhynchophylline on cell viability was detected with MTT.The protein levels of Bcl2,Bax,MMP3,MMP13,collagen II,Nrf2,and HO-1 were detected with Western Blotting.The levels of inflammatory cytokines PGE2,COX-1,TNF-α,and ROS were detected with ELISA.The level of MDA was used to detected with a kit.Results Compared to that in the control group,cell viability was repressed in IL-1βgroup(P<0.05).Compared to that in IL-1βgroup,cell viability increased in rhynchophylline(25μmol/L and 50μmol/L)groups(P<0.05).Compared to those in the control group,Bcl2 protein expression decreased and Bax protein expression elevated in IL-1βgroup(P<0.05).Compared to those in IL-1βgroup,Bcl2 protein expression increased in rhynchophylline(25μmol/L and 50μmol/L)groups and Bax protein expression was inhibited in rhynchophylline(50μmol/L)group(P<0.05).Compared to those in the control group,MMP13 and MMP3 protein levels were up-regulated and collagen II protein expression was down-regulated in IL-1βgroup(P<0.05).Compared to those in IL-1βgroup,MMP13 and MMP3 protein expression reduced in rhynchophylline(5μmol/L,25μmol/L and 50μmol/L)groups and collagen II protein expression was enhanced in rhynchophylline(25μmol/L and 50μmol/L)groups(P<0.05).Compared to those in the control group,the levels of PGE2,COX1,TNF-α,ROS,and MDA were promoted in IL-1βgroup(P<0.05).Compared to those in IL-1βgroup,the levels of PGE2,COX-1,TNF-α,ROS,and MDA were diminished in rhynchophylline(25μmol/L and 50μmol/L)groups(P<0.05).Compared to those in the control group,Nrf2 and HO-1 protein expression were reduced in IL-1βgroup(P<0.05).Compared to those in IL-1βgroup,Nrf2 and HO-1 protein expression increased in rhynchophylline(25μmol/L and 50μmol/L)groups(P<0.05).Conclusion Rhynchophylline inhibits IL-1β-induced oxidative stress and inflammatory damage in chondrocytes by activating Nrf2/HO-1 signaling pathway.
作者
王西彬
左瑞庭
王新立
李浩亮
刘汝银
WANG Xibin;ZUO Ruiting;WANG Xinli;LI Haoliang;LIU Ruyin(Department of Spine,He’nan Traditional Chinese Medicine Hospital,the Second Affiliated Hospital of Henan Chinese Medicine University,Zhengzhou 450002;Department of Rheumatology,He’nan Traditional Chinese Medicine Hospital,the Second Affiliated Hospital of Henan Chinese Medicine University,Zhengzhou 450002,China)
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2023年第7期966-970,986,共6页
Chinese Journal of Osteoporosis
基金
河南省中医药科学研究专项课题(2019ZYBJ18)。
