白介素6对人髓核细胞凋亡的影响  

Effect of interleukin-6 on apoptosis of human nucleus pulposus cells

作　　者：董振辉[1] 王德春[1] 

机构地区：[1]青岛大学医学院附属医院脊柱外科,青岛266003

出　　处：《中国矫形外科杂志》2010年第23期1976-1980,共5页Orthopedic Journal of China

基　　金：国家自然基金委课题项目(编号:30672132)

摘　　要：[目的]研究炎症因子IL-6对人髓核细胞凋亡的影响及可能机制。[方法]培养人髓核细胞,将细胞随即分为4组:IL-6刺激组;IL-6+P38MAPK阻断组;IL-6+JNK/SAPK阻断组及对照组。然后用AnnexinV/PI染色流式细胞仪和TUNEL法检测髓核细胞凋亡情况,免疫荧光检测P-P38MAPK和P-JNK/SAPK的表达及定位;West-ern印迹法检测P38MAPK、JNK/SAPK及各自活化形式,即P-P38MAPK和P-JNK/SAPK的表达。[结果]流式细胞仪和TUNEL法检测凋亡结果中,IL-6刺激组相对于对照组及各阻断组有着较高的凋亡细胞密度(P<0.01);免疫荧光结果显示IL-6刺激组P-P38MAPK和P-JNK/SAPK在细胞质和细胞核均有表达,各阻断组在细胞质和细胞核可见少量表达,与IL-6刺激组差异有显著统计学意义(P<0.01),对照组仅见极少量表达;Western印迹法结果显示P38MAPK,JNK/SAPK在各组髓核细胞内均有表达,但无活化形式,IL-6刺激组可见P-P38MAPK,P-JNK/SAPK表达,但相应阻断组无表达。[结论]外源性IL-6可通过P38MAPK和P-JNK/SAPK途径导致人髓核细胞凋亡。[Objective]To investigate the effects of IL-6 on apoptosis of human nucleus pulposus cells.[Methods]The human nucleus pulposus cells were cultured.The cells were divided into 4 groups at random：the group treated with IL-6,the group treated with IL-6＋20μmol/L SB203580,the group treated with IL-6 ＋20μmol/L SP600125 and the control group.The density of human nucleus pulposus cells of apoptosis was detected by flow cytometric assay and TUNEL method for each group.The expression and cellular localization of P-P38MAPK and P-JNK/SAPK were investigated by immunohistochemistry.The expression of total JNK /SAPK,P38MAPK and their phosphorylations were investigated by Western blotting.[Results]There was higher density of human nucleus pulposus cells of apoptosis in IL-6 group than that in the control group and those in inhibition groups（P0.01）.The expression of phosphorylations of P38MAPK and JNK /SAPK was increased after treatment with IL-6.The expression of phosphorylations of P38MAPK or JNK /SAPK was significantly inhibited after pretreatment with SB203580 or SP600125 compared with that in the IL-6 group in immunofluorescence analysis and Western blotting.Immunofluorescence analysis also demonstrated that P-P38MAPK and P-JNK/SAPK were expressed and distributed mainly in cytoplasm and nucleus,however,no P-P38MAPK and P-JNK/SAPK in the control group.[Conclusion]IL-6 induces the apoptosis of human nucleus pulposus cells via P38MAPK and JNK/SAPK pathway.

关 键 词：人髓核细胞 信号传导 炎症因子 P38MAPK JNK/SAPK 细胞凋亡 

分 类 号：R681.5[医药卫生—骨科学]