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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。
This article explored the effect of STCH (stress and chaperone) on dopaminergic neurons by different means. The expression of STCH was detected by RT-PCR. Single dopaminergic neurons were obtained by immuno-laser capture microdissection. The expression of dopaminergic neurons in different subregions of STCH was detected by RT-PCR. STCH Overexpression of pEGFP-N2 vector and pSuper-EGFP interference vector, transfected HEK293 and SH-SY5Y cell lines, detection of MPPT toxicity cells. The results showed that STCH was the lowest in the hippocampus and highest in the liver. The expression of dopaminergic neurons in the central gray matter area was undetectable in the substantia nigra. The STCH interference fragment was transfected into HEK293 cells with obvious cell death. To SH-SY5Y cells had no significant effect on the cell growth, but the cell morphology changed; SH-SY5Y cells over-expressing STCH compared with the control group were resistant to MPP + toxicity and inhibited the expression of caspase-12 expression. These results suggest that STCH has a potential protective effect on dopaminergic neurons, and its mechanism may be through inhibition of endoplasmic reticulum stress-induced apoptotic pathway. This result provides useful clues for finding therapeutic targets for Parkinson’s disease.