摘要
目的观察经6-羟基多巴胺(6-OHDA)诱导的MES23.5细胞神经元型一氧化氮合酶(nNOS)表达的变化,以及其在6-OHDA诱导的铁调节蛋白(IPR1)mRNA上调中的作用。方法应用10μmol/L的6-OHDA处理MES23.5细胞24h,制备帕金森病(PD)细胞模型,采用Western blots方法检测细胞中nNOS蛋白的表达,采用实时荧光定量PCR方法检测细胞中IRP1 mRNA的表达。结果 10μmol/L 6-OHDA处理的MES23.5细胞nNOS蛋白的表达水平较对照组明显增高,差异有显著性(t=16.72,P<0.001);6-OHDA处理组IRP1mRNA的表达水平较对照组明显增高,差异有显著性(F=6.07,q=4.84,P<0.0),而100μmol/L的nNOS抑制剂盐酸亚精胺预处理组IRP1表达较6-OHDA组明显降低,差异有显著性(q=4.02,P<0.05);盐酸亚精胺单独作用不影响IRP1mRNA的表达。结论 nNOS在6-OHDA制备的PD细胞模型中表达增高,且nNOS抑制剂盐酸亚精胺能明显抑制6-OHDA引起的IRP1上调。
Objective To observe the expression changes of nNOS induced by 6-OHDA as well as its role in 60HDA in- duced up-regulation of IRP1 in MES23.5 dopaminergic cells. Methods A Parkinson disease （PD） model was created using 10 μmol/L 6-OHDA to treat MES23.5 cells for 24 h. Western blot was used to detect the expression of nNOS in the cells and real- time PCR to detect the mRNA levels of IRPlin the model. Results The expression of nNOS in MES23.5 cells, treated with 10 μmol/L 6-OHDA, was higher than that in the control group, the difference being significant （t =16.72,P〈0.001）, and the ex pression of IRP1 mRNA also higher, treated same as that in the MES23.5 cells, than that in the control （F = 6.07 ,q=4.84, P 〈0.01）, while the expression of IRPI, pre-treated with 100 μmol/L spermidine hydrochloride, was lower than that in 60HNA group （q=4.02,P〈0.05）. Single action of spermidine hydrochloride did not impact IRP1 mRNA expression. Conclusion The expression of nNOS in Parkinson model created by 6-OHDA elevates, and nNOS inhibitor spermidine hydrochloride--can obviously inhibit the up-regulation of IRP1.
出处
《青岛大学医学院学报》
CAS
2016年第2期127-129,共3页
Acta Academiae Medicinae Qingdao Universitatis
基金
国家自然科学基金资助项目(81430024,31371081)
科技部国家重点基础研究发展计划973计划子课题资助项目(2011CB504102)
