大鼠髓核细胞体外衰老模型的建立

Establishment of the Senescence Model of Rat Nucleus Pulposus Cell in Vitro

目的:建立大鼠椎间盘髓核细胞体外衰老模型。方法:提取大鼠椎间盘髓核细胞,在完全培养基中培养,作为对照组;在对照组基础上加入终浓度为100μmol/L三丁基过氧化氢(t-BHP)培养2 h,构建髓核细胞体外衰老模型(衰老模型组)。采用Western印迹法检测两组髓核细胞的衰老相关指标微囊蛋白-1(caveolin-1)、β-半乳糖苷酶(SA-β-gal)的表达量,同时采用CCK-8细胞增殖实验检测两组的细胞活性。结果:与对照组比较,t-BHP作用2 h后,衰老模型组caveolin-1、SA-β-gal表达量明显升高,且髓核细胞增殖速率减慢,细胞活性明显降低。结论:采用t-BHP诱导的方法能成功构建髓核细胞体外衰老模型,caveolin-1在此过程中诱发了髓核细胞的衰老。

Objective：To establish a senescence model of rat intervertebral disc nucleus pulposus cells.Methods：The nucleus pulposus cells,which were extracted from rat intervertebral discs and cultured in complete medium,were set as control group.The senescence model of nucleus pulposus cells in vitro（senescence model group）was established by additional culture for two hours on the basis of control group,to which tert-butyl hydroperoxide（t-BHP）was added with a final concentration of 100μmol/L.The expression levels of senescence associated indexes,such as caveolin-1 and beta-galactosidase（SA-β-gal）,in two groups were assessed by Western blotting,while the cell viability in two groups was detected by Cell Counting Kit-8（CCK-8）proliferation assay.Results：After two hours of t-BHP function,the expression levels of caveolin-1and SA-β-gal in senescence model group were significantly higher than those in control group,while the nucleus pulposus cells proliferation was slower and the cell viability was much lower.Conclusions：The senescence model of nucleus pulposus cells in vitro can be established successfully with the induction of t-BHP,and caveolin-1 induces the senescence of nucleus pulposus cells during the process.