摘要
本文通过不同手段探讨了STCH(stress and chaperone)对多巴胺能神经元的作用。采用RT-PCR检测STCH的组织表达模式;采用免疫-激光捕获显微切割技术获得单一多巴胺能神经元,采用RT-PCR检测STCH在中脑不同亚区多巴胺能神经元的表达差异;分别构建STCH过表达pEGFP-N2载体和pSuper-EGFP干扰载体,转染HEK293和SH-SY5Y细胞株,检测细胞对MPP+毒性的反应。结果显示:STCH在海马表达最低,肝脏最高;在中央灰质区的多巴胺能神经元可检测到表达,在黑质区检测不到;将STCH干扰片段转染至HEK293细胞,细胞死亡明显;转染至SH-SY5Y细胞,对细胞生长无明显影响,但细胞形态发生改变;与对照组相比过表达STCH的SH-SY5Y细胞对MPP+毒性有抵抗作用,并能抑制MPP+处理细胞中caspase-12的表达。这些结果提示:STCH对多巴胺能神经元具有潜在的保护作用,其机制可能是通过抑制内质网应激诱导的凋亡途径而实现。该结果为寻找Parkinson病的治疗靶向提供了有益的线索。
We studied the protective role of STCH （ stress and chaperone） on dopaminergic neurons by different means： tissue expression pattern of STCH was examined by reverse-transcription polymerase chain reaction （RT-PCR） ; the differential STCH expression of dopaminergic （DA） neurons in different subregion of midbrain was examined with immuno-laser capture mierodisseetion combined with fluorescent quantitative RT-PCR; The pEGFP-N2 vector of recombinant ever-expression and interfering vectors of STCH-pSuper-EGFP were constructed respectively, and transfected into HEK293 and SH-SYSY cellular strain to observe their effects on MPP ^＋ toxicity. The results showed that hippocampus exhibited a lower expression, while liver exhibited the highest expression; no fluorescent signal was detected in DA neurons of substantia nigra, while DA neurons in central grey substance （CGS） showed expression; after STCH mRNA was knocked down in HEK293 cell line, the viability of the cells was significantly reduced, while when the same thing happened in SH-SY5 Y cell line, only the changed cell morphology was observed; in addition, STCH-transfected SH-SYSY cell showed resistance to MPP^＋ toxicity and the expression of caspase-12 was simuhaneously inhibited compared with control group. These results suggest that STCH have potential protective roles on DA neuron possibly by inhibition of endoplasmic reticular stress-induced apoptosis pathway. Those data thus provide useful clues to find new therapeutic target for Parkinson＇s disease.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2008年第6期592-596,共5页
Chinese Journal of Neuroanatomy
基金
上海市卫生局科技发展基金(No.054091)
上海市自然科学基金(No.07ZR14122)资助项目
