6-羟多巴胺对SH-SY5Y细胞信号转导和转录激活因子1和3磷酸化水平的影响  

Effect of 6-hydroxydopamine on the levels of phosphorylation of signal transduction and transcription factor 1 and 3:experiment with dopaminergic neurons

作　　者：王莉莉[1] 陈彪[1] 徐胜利[1] 

机构地区：[1]首都医科大学宣武医院神经内科,北京100053

出　　处：《中华医学杂志》2007年第15期1048-1052,共5页National Medical Journal of China

基　　金：国家自然科学基金重点项目(30430280);北京市自然科学基金重点项目(7031002)

摘　　要：目的观察6-羟多巴胺（6-OHDA）对SH-SYSY细胞信号转导和转录激活因子STATl和STAT3磷酸化水平的影响，探讨STAT在6-OHDA神经毒性机制中的作用。方法用不同浓度的6-OHDA处理SH-SY5Y细胞制备细胞损伤模型，用^3H标记胸腺嘧啶（^3H-TdR）掺入实验和噻唑蓝（MTT）方法检测细胞生存率。用Western印迹检测细胞STAT1和STAT3磷酸化水平的改变。结果6-OHDA以浓度和时间依赖性的方式引起SH-SY5Y细胞生存率下降、STAT3磷酸化水平降低而对STAT1磷酸化水平无影响。50—400μmol／L6-OHDA处理细胞24h，细胞生存率下降为对照组的8．1％-81．9％。100μmol／L6-OHDA处理细胞0．5～48h引起STAT3磷酸化水平明显下降，条带相对密度为0．747±0．011～0．238±0．007。50～200μmol／L的6-OHDA处理细胞2h也引起了STAT3磷酸化的明显下降，条带相对密度为0．895±0．003～0．633±0．002。以上结果和对照组相比均有统计学意义（均P〈0．05）。结论6-OHDA可能通过引起细胞STAT3磷酸化水平下降而导致细胞损害。Objective To observe the effects of 6-hydroxydopamine （6-OHDA） on the levels of phosphorylation of signal transducer and activator of transcription （STAT） 1 and STAT3, and the role of STASD in the neurotoxicity of 6-OHDA on dopaminergic neurons so as to help study the treatment of Parkinson＇s disease. Methods Dopaminergic neurons, human neuroblastoma cells of the line SH-SYSY were cultured and were treated with 6-OHDA of the concentrations of 50, 100, 200, and 400 μmol/L respectively. SH-SYSY cells not treated with 6-OHDA were used as control group. MTT method was used to observe the cell viability rate 24, 48, and 72 h later. 3H-thymidine deoxyribose （TdR） was added into the culture fluid 12, 36, or 60 h after the treatment of 6-OHDA for 12 h, then the cell viability rate was observed. Western blotting was used to detect the levels of STAT1 and STAT3 phosphorylation. Results Twenty-four hours after the treatment of 50-400 μmol/1 6-OHDA the cell viability rates decreased to 8. 1% - 81.9% that of the control cells （ all P 〈 0. 05 or P 〈 0.01 ） , e. g. , 100 μmol/L 6-OHDA decreased the cell viability rate to 50% that of the control cells （P 〈 0.01 ）. 3H-TdR proliferation assay showed that the cell viability rate decreased with the increase of the 6-OHDA concentration and time of treatment too. Western blotting showed that 0. 5-24 h after the treatment with 100 μmol/L 6-OHDA, the STAT3 phosphorylation level was decreased obviously with the comparative band density of 0. 747 ± 0. 011-0. 238 ± 0. 007, and 2 hours after the treatment with 50-200 μmol/L 6-OHDA the comparative band density of STAT3 phosphorylation was 0. 895 ± 0. 003-0. 633 ± 0. 002 （ all P 〈 0. 05 ） . However, the level of STAT1 phosphorylation was not influenced by the treatment of 6-OHDA of the concentration of 100 mol/L at different time points. Conclusions Decreased STAT3 phosphorylation may be involved in the neurotoxicity mechanisms of 6-OHDA on dopaminergic neurons.

关 键 词：帕金森病 羟多巴胺 信号传导 

分 类 号：R363[医药卫生—病理学]