Fas配体对椎间盘细胞经典凋亡通路中关键因子的影响  

Effect of Fas ligand on the key factors in apoptotic pathway of intervertebral disc cells

作　　者：韩敦富[1] 解立俊[2] 吴海燕[1] 胡法梅 尹荷珊 

机构地区：[1]山东省淄博市中心医院微创脊柱外科,淄博255036 [2]山东省淄博市中心医院麻醉科,淄博255036 [3]山东省淄博市中心医院皮肤科,淄博255036

出　　处：《安徽医科大学学报》2013年第12期1441-1444,共4页Acta Universitatis Medicinalis Anhui

基　　金：山东省医药卫生科技发展计划项目(编号:2011WSA03016)

摘　　要：目的明确Fas配体(FasL)介导的大鼠椎间盘细胞的确切凋亡途径。方法分离、培养大鼠腰椎间盘髓核细胞,分别与含不同浓度FasL(0、5、10、20、50 ng/ml)的1%胎牛血清(FBS)培养基共培养24 h,分别检测Fas,Caspase-8、9、3及Bid mRNA的表达水平,Caspase-8、9、3的酶活性以及线粒体膜电势的变化。结果髓核细胞与不同浓度FasL共培养24 h后,经Hoechst 33258染色均可观察到细胞凋亡,Caspase-8、3及Bid mRNA的表达水平,Caspase-8、3的酶活性均随FasL浓度的升高而升高;50 ng/ml的FasL可明显增强Caspase-9 mRNA的表达、Caspase-9的酶活性和线粒体膜电势。结论 FasL所诱发的椎间盘细胞凋亡主要通过膜途径进行,大剂量时可以激活线粒体途径。Objective To investigate the apoptotic effect and identify the apoptotic pathway of Fas ligand （FasL） on rat intervertebral disc cells in vitro. Methods ① Rat nucleus pulposus ceils were cocuhured with different dose of recombinant rat FasL （0, 5, 10, 20, 50） in 1% FBS medium for 24 h, then the expression levels of Fas, Caspase-8, Caspase-9, Caspase-3 and Bid mRNA were detected by RT-PCR, the enzyme activity of Caspase-8, Caspase-9, Caspase-3 was evaluated with enzyme activity detection Kit by enzyme appearance, membrane potential change of mitochondria was determined using fluorescence microscope. Results Nucleus pulposus cells with different concentrations FasL training after 24 h, cell apoptosis was all observed by Hoechst 33258 dyeing. The levels of Caspase-8, Caspase-3 and Bid mRNA were detected by RT-PCR, the enzymatic activity of Caspase-8 and Caspase- 3 rised respectively with the dose of recombinant rat FasL. The mRNA expression level and enzymatic activity of Caspase-9, and membrane potential change of mitochondria were increased by 50 mg/ml FasL. Conclusion Membrane apoptotic pathway is the main road of rat NP cells＇ apoptosis induced by FasL. And mitochondrium apoptotic pathway can be activated by a higher dose of FasL.

关 键 词：椎间盘细胞 凋亡 凋亡途径 FAS配体 

分 类 号：R318[医药卫生—生物医学工程]