川芎嗪抑制NF-κB P65磷酸化对LPS诱导的骨关节炎软骨细胞凋亡和炎症反应的调节作用  

Ligustrazine regulates LPS-induced apoptosis and inflammatory response of osteoarthritis chondrocytes via inhibiting phosphorylation of NF-κB P65

作　　者：朱海泉 刘子敏 孟祥圣 孙晓[1] 王黎明[2] ZHU Hai-Quan;LIU Zi-Min;MENG Xiang-Sheng;SUN Xiao;WANG Li-Ming(Department of Emergency Surgery,Lianyungang Clinical Medicine Hospital of Nanjing Medical University,Lianyungang 222002,China)

机构地区：[1]南京医科大学连云港临床医学院急诊外科,连云港222002 [2]南京医科大学附属南京医院骨科,连云港222002

出　　处：《中国免疫学杂志》2019年第2期181-185,共5页Chinese Journal of Immunology

基　　金：江苏省青年基金资助(BK20160423)

摘　　要：目的:探讨川芎嗪对LPS诱导的骨关节炎软骨细胞凋亡和炎症反应的影响及机制。方法:LPS诱导骨关节炎模型。软骨细胞分为4个组:对照组;川芎嗪(20μmol/L)组; LPS(100 ng/ml)组;川芎嗪(20μmol/L)+LPS(100 ng/ml)组。Hoechst33258染色检测细胞凋亡。ELISA检测一氧化氮(Nitric oxide,NO)、肿瘤坏死因子-α(TNF-α)和白细胞介素(IL)-6水平。蛋白印迹检测Ⅱ型胶原(collagenⅡ)、聚集蛋白聚糖(aggrecan)和基质金属蛋白酶-13(MMP-13)、NF-κB P65和p-NF-κB P65蛋白水平。结果:川芎嗪(20μmol/L)可改善LPS诱导的骨关节炎软骨细胞形态异常及细胞数量的减少。LPS组细胞凋亡率明显高于对照组(P<0. 05)。川芎嗪可降低LPS诱导的细胞凋亡率的升高(P<0. 05)。与对照组相比,LPS组Ⅱ型胶原和聚集蛋白聚糖表达明显降低,MMP-13表达明显升高(P<0. 05)。川芎嗪可抑制LPS诱导的Ⅱ型胶原和聚集蛋白聚糖表达水平的下降及MMP-13表达水平的上升(P<0. 05)。LPS组NO、TNF-α和IL-6水平明显高于对照组(P<0. 05)。LPS+川芎嗪组NO、TNF-α和IL-6水平显著低于LPS组(P<0. 05)。与对照组相比,LPS组p-P65/P65比值明显升高(P<0. 05)。川芎嗪可减弱LPS诱导的p-P65/P65比值的上升(P<0. 05)。结论:川芎嗪通过抑制NF-κB P65磷酸化减轻LPS诱导的骨关节炎软骨细胞凋亡和炎症反应。Objective:To explore the effect of ligustrazine on the LPS-induced apoptosis and inflammatory response of osteoarthritis chondrocytes.Methods:Osteoarthritis model was induced by LPS.Chondrocytes were divided into four group:control group,ligustrazine(20μmol/L)group,LPS(100 ng/ml)group and ligustrazine(20μmol/L)+LPS(100 ng/ml)group.Apoptosis was measured by Hoechst33258 staining.The levels of nitric oxide(NO),tumor necrosis factorα(TNF-α)and interleukin(IL)-6 were detected by ELISA.The protein levels of collagenⅡ,aggrecan,matrix metalloproteinase 13(MMP-13),NF-κB P65 and p-NF-κB P65 were tested by Western blot.Results:The LPS-induced abnormal cell morphology and decreased number of cells were ameliorated by ligustrazine(20μmol/L).The apoptosis in LPS group was higher than control group(P<0.05).The LPS-induced enhancive apoptosis was reduced by ligustrazine(P<0.05).Compared with control group,the expression of collagenⅡand aggrecan was alleviated with increased expression of MMP-13(P<0.05).The LPS-induced declined expression of collagenⅡand aggrecan and elevated expression of MMP-13 was inhibited by ligustrazine(P<0.05).The levels of NO,TNF-αand IL-6 in LPS group were higher than control group(P<0.05).The levels of NO,TNF-αand IL-6 in LPS+ligustrazine group were lower than LPS group(P<0.05).Compared with control group,the rate of p-P65/P65 in LPPS group was enhanced(P<0.05).The LPS-indiced increased rate of p-P65/P65 was decreased by ligustrazine(P<0.05).Conclusion:Ligustrazine alleviates the LPS-induced apoptosis and inflammatory response of osteoarthritis chondrocytes via inhibiting phosphorylation of NF-κB P65.

关 键 词：川芎嗪 骨关节炎 细胞凋亡 炎症 NF-ΚB P65 

分 类 号：R684.3[医药卫生—骨科学]