H2O2氧化应激诱导兔椎间盘髓核细胞衰老的研究
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作者Author单位AddressE-Mail
谢健 XIE Jian 浙江中医药大学, 浙江 杭州 310053  
童培建 TONG Pei-jian 浙江省中医院骨科, 浙江 杭州 310006 Traditional Chinese Medicine Hospital of Zhejiang Province, Hangzhou 310006, Zhejian, Chinese  
单乐天 SHAN Le-tian 浙江中医药大学, 浙江 杭州 310053  
吴承亮 WU Cheng-liang 浙江中医药大学, 浙江 杭州 310053 wu.cl@163.com 
期刊信息:《中国骨伤》2013年,第26卷,第4期,第332-335页
DOI:10.3969/j.issn.1003-0034.2013.04.017
基金项目:浙江省自然科学基金(编号:Y20091189);浙江省医药卫生科技计划项目(编号:2010KYA142)
中文摘要:

目的: 研究H2O2不同浓度对兔椎间盘髓核细胞形态、活力、增值、周期等的影响。

方法: 新西兰大白兔(2~3 kg,雌)10只, 无菌条件下酶消化法分离髓核细胞。含15%FBS的DMEM/F12(1:1)培养液培养, 细胞90%融合后传第1代。按照H2O2不同浓度(0 μmol/L、130 μmol/L、216 μmol/L、360 μmol/L、600 μmol/L、1 000 μmol/L)分组, 0 μmol/L H2O2为空白对照组。在细胞对数生长期, 不同浓度H2O2处理1 h后原培养液继续培养48 h,通过检测, 分析比较各组髓核细胞与空白对照组间形态、活力、增值、周期的差异性。

结果: 与空白对照组比较, 当H2O2浓度为130 μmol/L、216 μmol/L时, 髓核细胞生物学特性未见显着性改变(P>0.05);当H2O2浓度为360 μmol/L、600 μmol/L、1 000 μmol/L时, 髓核细胞出现衰老改变:细胞质减少并出现空泡、增值减慢、衰老相关-β-半乳糖苷酶蓝染率增加、细胞周期阻滞于G1期而进入S期减少(P<0.05).随着H2O2浓度的增高, 衰老程度不断升高。

结论: 一定浓度的H2O2能够导致髓核细胞提前衰老, 引起髓核细胞生物学特性的改变。
【关键词】椎间盘  细胞衰老  过氧化氢  
 
Reseach of oxidative stress induces aging in rabbit intervertebral disc nucleus pulposus cells injured by H2O2
ABSTRACT  

Objective: To study the effect of H2O2 on the morphological pattern,vitality,proliferation,cycle period of rabbit intervertebral disc nucleus pulposus cells.

Methods: Ten New Zealand white rabbits (2 to 3 kg,female) were used for isolating nucleus pulposus cells under sterilized condition. The culture solution with 15% FBS and DMEM/F12 (1∶1) was applied for cell cultivation. After 90% cell fusion,the first generation was obtain and stimulated by H2O2 with different concentrations of 0 μmol/L (control group),130 μmol/L,216 μmol/L,360 μmol/L,600 μmol/L,and 1000 μmol/L.

Results: Compared with the control group,there was little difference of the biological property (P>0.05) in 130 μmol/L and 216 μmol/L H2O2-treated groups. When the concentration of H2O2 attained 360 μmol/L,600 μmol/L,and 1 000 μmol/L,the cells suffered aging,with increased cell vacuoles,decreased proliferation,and aging∶related increase of β-galactosidase dyeing. The cell cycle of many nucleus pulposus cells was blocked in G1 stage other than entering S stage. With increasing H2O2 concentrations,the aging degree was increased.

Conclusion: A certain concentration of H2O2 could induce early aging of nucleus pulposus cells,resulting in biological abnormalities of these cells.
KEY WORDS  Intervertebral disk  Cell aging  Hydrogen peroxide  Rabbits
 
引用本文,请按以下格式著录参考文献:
中文格式:谢健,童培建,单乐天,吴承亮.H2O2氧化应激诱导兔椎间盘髓核细胞衰老的研究[J].中国骨伤,2013,26(4):332~335
英文格式:XIE Jian,TONG Pei-jian,SHAN Le-tian,WU Cheng-liang.Reseach of oxidative stress induces aging in rabbit intervertebral disc nucleus pulposus cells injured by H2O2[J].zhongguo gu shang / China J Orthop Trauma ,2013,26(4):332~335
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