N-乙酰半胱氨酸对骨关节炎软骨细胞氧化应激保护作用的研究  

Protective Effect of N-Acetylcysteine on Oxidative Stress in Osteoarthritic Chondrocytes

作　　者：陈劲宇 洪鸿翔 袁锟[1] 朱新辉[1] 王飞[1] 周小刚 范艳艳 CHEN Jin-yu;HONG Hong-xiang;YUAN Kun;ZHU Xin-hui;WANG Fei;ZHOU Xiao-gang;FAN Yan-yan(Department of Orthopaedics,the First People's Hospital of Nantong City,Nantong 226000,China;Department of Orthopaedics,Nantong Hospital of Traditional Chinese Medicine,Nantong 226001,China)

机构地区：[1]江苏省南通市第一人民医院骨科,江苏南通226000 [2]江苏省南通市中医院骨科,江苏南通226001

出　　处：《药物生物技术》2019年第6期505-508,共4页Pharmaceutical Biotechnology

摘　　要：文章围绕N-乙酰半胱氨酸对骨关节炎软骨细胞氧化应激保护作用展开研究。筛选2017年1月~2018年12月前往南通第一人民医院骨科接受膝关节置换术治疗的10例骨关节炎患者,另选同期5例因外伤致截肢患者,在征得患者知情同意下采集膝关节软骨,提取软骨细胞后将骨关节炎软骨细胞以及正常软骨细胞各一分为二,不含N-乙酰半胱氨酸培养基培养的分别设为骨关节炎Ⅰ组和正常软骨Ⅰ组,包含N-乙酰半胱氨酸培养基培养的骨关节炎Ⅱ组和正常软骨Ⅱ组,体外培养24 h后对氧化应激指标(丙二醛、超氧化物歧化酶、一氧化氮、一氧化氮合成酶)进行测定,利用实时定量荧光PCR对细胞核因子E2相关因子mRNA及其蛋白表达水平进行测定,Western blot法测定细胞核因子E2相关因子mRNA及其蛋白表达水平,荧光共聚焦显微镜下观察核内细胞核因子E2相关因子活性。体外培养前骨关节炎Ⅰ组和Ⅱ组、正常软骨Ⅰ组和Ⅱ组氧化应激指标、细胞核因子E2相关因子mRNA及其蛋白表达水平、核内细胞核因子E2相关因子活性无统计学差异(P> 0. 05)。体外培养24 h后骨关节炎Ⅰ组和Ⅱ组丙二醛、核内细胞核因子E2相关因子活性较体外培养前下降,超氧化物歧化酶、一氧化氮、一氧化氮合成酶、细胞核因子E2相关因子mRNA及其蛋白表达水平较体外培养前上升,存在统计学差异(P <0. 05),正常软骨Ⅰ组和Ⅱ组与体外培养前无统计学差异(P> 0. 05)。体外培养24 h后,骨关节炎Ⅰ组和Ⅱ组相比,存在统计学差异(P <0. 05),正常软骨Ⅰ组和Ⅱ组相比,无统计学差异(P> 0. 05),骨关节炎组与正常软骨组相比,存在统计学差异(P <0. 05)。N-乙酰半胱氨酸能够抑制骨关节炎软骨细胞氧化应激反应,激活细胞核因子E2相关因子以降低其受到的损伤。To study the protective effect of N-acetylcysteine on oxidative stress of osteoarthritic chondrocytes,from January 2017 to December 2018,10 patients with osteoarthritis who underwent knee arthroplasty in the Department of Orthopaedics,Nantong First People’s Hospital were selected. Another 5 patients with amputation due to trauma were enrolled with informed consent. The knee articular cartilage was collected,and the osteoarthritic chondrocytes and normal chondrocytes were divided into two groups,and the N-acetylcysteine-free medium was cultured as osteoarthritis group I and normal cartilage I group,including osteoarthritis group II and normal cartilage group II cultured in N-acetylcysteine medium,oxidative stress index( malondialdehyde,superoxide dismutase,nitric oxide,one after 24 hours culture in vitro). Nitric oxide synthase was determined by real-time quantitative fluorescent PCR. The expression of nuclear factor E2 related factor mRNA and its protein was determined by Western blot. Nuclear nuclear factor E2-related factor activity in the nucleus was observed by FCFM. There were no significant differences in oxidative stress index,nuclear factor E2 related factor mRNA and protein expression,and nuclear nuclear factor E2 related factor activity between group I and group II,normal cartilage group I and group II.( P > 0. 05),the activity of malondialdehyde and nuclear nuclear factor E2 related factors in osteoarthritis group I and group II decreased after 24 hours in vitro,superoxide dismutase,nitric oxide,nitric oxide synthase The expression of nuclear factor E2 related factor mRNA and protein increased compared with that in vitro( P < 0. 05),there was no significant difference between normal cartilage group I and group II before culture( P > 0. 05). There was a statistically significant difference between the arthritis group I and the group II( P < 0. 05). There was no significant difference between the normal cartilage group I and the group II( P > 0. 05). Between the osteoarthritis group and the normal cart

关 键 词：骨关节炎 软骨细胞 N-乙酰半胱氨酸 氧化应激 细胞核因子E2相关因子 丙二醛 超氧化物歧化酶 

分 类 号：R735.7[医药卫生—肿瘤]