丙戊酸钠对6-OHDA诱导的SH-SY5Y细胞凋亡的保护作用及其机制

目的:研究丙戊酸(VPA)对6-OHDA引起的SH-SY5Y细胞凋亡的抑制作用及其机制。方法:采用光镜观察细胞形态,监测其存活状态,Hoechst33342荧光染色检测细胞凋亡。采用Western blotting法检测细胞中抗凋亡蛋白Bcl-2的表达。结果:形态学观察发现,6-OHDA处理SH-SY5Y细胞使大部分细胞胞体皱缩,突起缩短、消失或断裂,细胞存活率显著下降;而VPA(1mmol/L)预处理细胞24h,可提高细胞存活率。Hoechst33258荧光染色后出现染色质固缩等细胞凋亡的特征性改变,而用VPA预处理SH-SY5Y细胞可明显减少SH-SY5Y细胞凋亡。Western Blotting分析证实,6-OHDA抑制Bcl-2蛋白的表达,而VPA预处理Bcl-2蛋白含量明显上调。结论:VPA能抑制6-OHDA诱导的SH-SY5Y细胞凋亡,Bcl-2可能是VPA神经保护作用的一个重要靶点。

Valproate provides a protective effect against 6-OHDA induced apoptosis in SH-SY5Y cells

Objectives: To study the inhibiting effect of VPA on 6-OHDA induced apoptosis in SH-SY5Y cells. Methods: The cell morphology and survival were observed by light microscope. Hoechst 33342 staining assay was applied to detect apoptosis . The anti-apoptosis protein Bcl-2 was determined with Western blot. Results: Light microscope revealed that most cell bodies shrank and neurites became shorter or broken, even disappeared and the cell survival decreased after 6-OHDA treatment. However, VPA would increase the cell survival and extend the neurites. Hoechst 33342 staining showed that apoptosist features including chromosome pyknosis appeared in SH-SY5Y cells induced by 6-OHDA. While it attenuated by pretreatment with VPA,Western blotting analysis indicated that 6-OHDA decreased bcl-2 expression and VPA could up-regulate the level of Bcl-2. Conclusion: VPA could prevent the apoptosis of SH-SY5Y cells caused by 6-OHDA. Bcl-2 might be the principal target for the neuroprotection exerted by VPA.