瘦素减轻6-羟基多巴胺诱导的SH-SY5Y细胞损伤  

Leptin alleviates damage of 6-OHDA-induced SH-SY5Y cells

作　　者：韩铭[1] 邓子辉[1] 薛辉[1] 李永明[1] 颜光涛[1] 

机构地区：[1]解放军总医院基础所生化室,北京100853

出　　处：《解放军医学院学报》2014年第1期70-73,77,共5页Academic Journal of Chinese PLA Medical School

基　　金：科技基础性工作专项项目(2011FY130100);国家科技支撑计划项目(2012BAK25B01)~~

摘　　要：目的探讨瘦素对6-羟基多巴胺(6-hydroxydopamine,6-OHDA)所诱导的人神经母细胞瘤细胞(SH-SY5Y)凋亡的抑制作用。方法建立6-OHDA诱导的SH-SY5Y细胞损伤模型,用瘦素进行干预,采用倒置显微镜观察细胞形态,MTT法检测细胞活性,LDH法检测细胞损伤,台盼蓝检测细胞死亡率,流式细胞术检测细胞凋亡率,并用免疫荧光检测active caspase-3的荧光强度,Western blot检测Bcl-2和Bax的表达水平。结果成功建立了6-OHDA诱导的SH-SY5Y细胞损伤模型。在模型的基础上,瘦素干预后,细胞凋亡率降低约15%,细胞存活率提高约14%,细胞损伤降低率和死亡率,active caspase-3的浓度下降,Bcl-2/Bax的比值显著提高(P<0.05)。结论瘦素对6-OHDA诱导的SH-SY5Y细胞凋亡具有一定的抑制作用,这对瘦素可能参与帕金森病的保护作用提供了证据。Objective To study the inhibitory effect of leptin on 6-OHDA-induced apoptosis of SH-SY5Y cells. Methods A 6-OHDA-induced SH-SY5Y cells damage model was established and treated with leptin. Morphology of SH-SY5Y cells was observed under inverted microscope, viability of SH-SY5Y cells was assayed by MTT, damage of SH-SY5Y cells was detected by LDH, death rate of SH-SY5Y ceils was tested with trypan blue staining, apoptosis of SH-SY5Y cells was detected by flow cytometry, fluorescence intensity of active caspase-3 was measured by immunofluorescence and expression levels of Bcl-2 and Bax were measured by Western blot. Results The 6-OHDA-induced SH-SY5Y cells damage model was successfully established. The apoptosis of SH-SY5Y cells was about 15% lower, the survival rate of SH-SY5Y cells was about 14% higher, the damage and death of SH-SY5Y cells and the active caspase-3 concentration were higher, and the Bcl-2/Bax ratio was significantly lower after leptin treatment than before leptin treatment （P 〈 0.05）. Conclusion Leptin can inhibit 6-OHDA -induced apoptosis of SH-SY5Y cells, which provides the evidence for leptin in prevention of Parkinson＇s disease.

关 键 词：6-羟基多巴胺 SH—SY5Y细胞 细胞损伤 瘦素 

分 类 号：R34[医药卫生—基础医学]