p38MAPK信号转导通路在氧化应激诱导兔髓核细胞衰老过程中的作用研究  

Role of p38MAPK Signaling Pathway in the Oxidative Stress-induced Aging Process of Rabbit Nucleus Pulposus Cells

作　　者：吴承亮[1] 谢健[1] 廖菲[1] 

机构地区：[1]浙江中医药大学,杭州310053

出　　处：《浙江中医药大学学报》2013年第5期489-492,共4页Journal of Zhejiang Chinese Medical University

基　　金：浙江省医药卫生科技计划项目(2010KYA142);浙江省自然科学基金项目(Y2091189);浙江中医药科技计划项目(2009CA001)~~

摘　　要：[目的]研究p38MAPK信号转导通路在氧化应激诱导兔髓核细胞衰老过程中的作用。[方法]2～3月龄新西兰雌性大白兔10只,无菌条件下酶消化法分离其髓核细胞。原代培养兔髓核细胞传第一代,将生长良好的髓核细胞制成细胞悬液,随机分为空白组、模型组和阻断组。空白组为正常髓核细胞;模型组以浓度为0.36mmo.lL-1过氧化氢作用1h,造成髓核细胞的氧化应激模型;阻断组以p38MAPK抑制剂预处理髓核细胞30min后,再按照模型组造成氧化应激模型。各组经上述处理后继续培养48h,进行髓核细胞形态学观察,同时检测髓核细胞增殖率、β-半乳糖苷酶(SA-β-gal)活性及细胞周期,最后进行组间比较分析。[结果]各组髓核细胞形态未出现显著变化,但模型组和阻断组的细胞分布密度较稀疏、并有典型衰老改变,主要表现为:细胞增殖减慢、衰老相关β-半乳糖苷酶蓝染率增加、细胞周期阻滞于G1期等(与空白组相比P<0.05)。其中模型组衰老程度最为严重,阻断组则抑制了部分衰老改变(与模型组相比P<0.05)。[结论]p38MAPK信号转导通路是介导氧化应激诱导兔髓核细胞衰老和凋亡过程的关键通路之一。[Objective]To investigate the role of p38MAPK signaling pathway in the oxidative stress induced aging process of rabbit nucleus pulposus cells.[Methods]10 New Zealand white female rabbits（2～3 month old） were used,and the nucleus pulposus cells were separated by enzyme digestion under the aseptic condition.Primary rabbit nucleus pulposus cells were cultured to obtain the first generation of the cells.The well-grown cells were prepared as cell suspension and divided into normal group,model group（H2O2 treated） and inhibition group（SB203580＋H2O2 treated）.The normal group was the blank control with no treatment,the model group was treated with H2O2 at 0.36 mmol·L-1 for 1h,and the inhibition group was treated with H2O2 following p38MAPK inhibitor（SB203580） pretreatment.All groups were thereafter cultivated for 48h and the cell morphological observation,cell proliferation assay,β-galactosidase（SA-β-gal） activity assay,and cell cycle assessment were performed for comparison among the different groups.[Results] There was little morphological difference among the three groups,whereas the cell density decrease with typical aging abnormality was found in the model group and inhibition group.When compared with the normal group,a decrease of cell proliferation,increase of β-galactosidase dyeing rate,and blocking of cell cycle in G1 stage were found in the model group and inhibition group（P＆lt;0.05）.The aging degree of the model group was more severe than that of the inhibition group（P＆lt;0.05）.[Conclusion] p38MAPK signaling pathway is an important pathway which can mediate the oxidative stress induced aging and apoptosis of rabbit nucleus pulposus cells.

关 键 词：髓核细胞 氧化应激 信号转导 P38MAPK 细胞衰老 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]