组蛋白去乙酰化酶6对帕金森病细胞模型中α-突触核蛋白阳性包涵体的作用

Effects of histone deacetylase 6 on α-synuclein-positive inclusion bodies in the cell model of Parkinson's disease

目的研究帕金森病(PD)细胞模型中组蛋白去乙酰化酶6(HDAC6)的表达水平及其对α-突触核蛋白阳性包涵体的影响。方法使用Lipofectamin 2000构建稳定过表达野生型α-突触核蛋白的人神经母细胞瘤SK-N-SH细胞,并加入蛋白酶体抑制剂lactacystin制作α-突触核蛋白异常聚集的PD模型;采用Western blotting检测HDAC6的表达水平;使用HDAC6特异性抑制剂tubacin处理后,免疫荧光染色检测α-突触核蛋白阳性的包涵体水平。结果 Lactacystin处理细胞的HDAC6表达水平较对照细胞明显升高,且α-突触核蛋白阳性的包涵体增多,而经tubacin处理后包涵体减少,差异均有统计学意义(P<0.05)。结论在蛋白酶体抑制剂制作的PD细胞模型中,抑制升高的HDAC6可使α-突触核蛋白阳性的包涵体水平降低;其机制可能与HDAC6参与α-突触核蛋白从寡聚体向包涵体形式的转化从而起保护作用有关。

Objective To investigate the expression of histone deacetylase 6（HDAC6） and its effects on asynuclein-positive inclusion bodies in the cell model of Parkinson＇s disease（PD）.Methods The human neuroblastoma cells SK-N-SH with stable over-expressed wild type a-synuclein were constructed via Lipofectamin2000.The PD model with abnormal aggregation of a-synuclein was established by adding the proteasome inhibitor lactacystin.The expression of HDAC6 was detected by Western blotting.After being treated with HDAC6 specific inhibitor tubacin,the level of a-synuclein-positive inclusion bodies was detected by immunofluorescence staining.Results Compared with the controls,the expression of HDAC6 of lactacystintreated cells was significant higher and a-synuclein-positive inclusion bodies were more.Treatment by tubacin reduced the amount of inclusion bodies and the difference was statistically significant（P〈0.05）.Conclusion In the PD cell model established by proteasome inhibitor,inhibition of increased HDAC6 can reduce the amount of a-synuclein-positive inclusion bodies,which may be relevant to the involvement of HDAC6 in the conversion of a-synuclein from oligomers to inclusion bodies,so as to play a role of protection.