Kao Xi-bin, Gao Yan, Chen Jing-hong, Chen Qun, Wang Zhi-lun, Wang Zhou
Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi'an Jiaotong University, Shanxi Province 710061, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2013 Apr;31(4):271-5.
To study the role of c-jun N-terminal kinase (JNK) signaling pathway in chondrocyte apoptosis induced by nitric oxide (NO) using NO donor sodium nitroprusside (SNP) and JNK inhibitor SP600125.
Articular chondrocytes were separated from New Zealand rabbits aged 3 weeks by mechanical digestion and enzyme digestion and identified by toluidine blue staining, and then the chondrocytes were treated with SNP and SP600125 for 24 h. The cell apoptosis was evaluated by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL), and the expression levels of nuclear factor-kappa B (NF-κB) p65 and p53 were measured by western blot.
Compared with those in control group, the early apoptotic rate of SNP-treated chondrocytes increased as the concentration of SNProse, exhibiting a concentration dependency (P < 0.05), and the expression levels of NF-κB p65 and p53 also increased (P < 0.05); JNK inhibitor SP600125 inhibited these increases (P < 0.05).
JNK signaling pathway plays an important role in NO-induced chondrocyte apoptosis. JNK inhibitor SP600125 can reduce NO-induced apoptosis and expression of NF-κB p65 and p53 in articular chondrocytes of rabbits in a concentration-dependent manner.
采用一氧化氮（NO）供体硝普钠（SNP）及JNK抑制剂SP600125，研究c-Jun氨基末端激酶（JNK）信号通路在NO诱导的软骨细胞凋亡中的作用。
采用机械消化和酶消化法从3周龄新西兰兔分离关节软骨细胞，经甲苯胺蓝染色鉴定后，用SNP和SP600125处理软骨细胞24小时。采用膜联蛋白V-异硫氰酸荧光素（FITC）/碘化丙啶（PI）流式细胞术和末端脱氧核苷酸转移酶（TdT）介导的dUTP-生物素缺口末端标记法（TUNEL）评估细胞凋亡情况，采用蛋白质免疫印迹法检测核因子κB（NF-κB）p65和p53的表达水平。
与对照组相比，SNP处理的软骨细胞早期凋亡率随SNP浓度升高而增加，呈浓度依赖性（P<0.05），NF-κB p65和p53的表达水平也升高（P<0.05）；JNK抑制剂SP600125可抑制上述升高（P<0.05）。
JNK信号通路在NO诱导的软骨细胞凋亡中起重要作用。JNK抑制剂SP600125可呈浓度依赖性降低NO诱导的兔关节软骨细胞凋亡及NF-κB p65和p53的表达。
