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通过简单染料-脂质复合物检测发现,用鱼藤酮灌注导致偏侧帕金森病的大鼠纹状体中磷脂水平降低。

Attenuated levels of phospholipids in the striatum of rats infused with rotenone causing hemiparkinsonism as detected by simple dye-lipid complex.

作者信息

Gopi Margabandhu, Vanisree Arambakkam Janardhanam

机构信息

Department of Biochemistry, University of Madras, Guindy Campus, Chennai 600 025, Tamilnadu, India.

出版信息

IBRO Rep. 2017 Jun 13;3:1-8. doi: 10.1016/j.ibror.2017.06.001. eCollection 2017 Dec.

Abstract

Parkinson's disease (PD), a progressive neurodegeneration, is characterized by loss of dopaminergic neurons in the substantia nigra (SN) and loss of motor co-ordination. Impaired metabolism of major lipids such as phospholipids which play regulatory roles in cellular functions and signaling has been implicated in the pathology of PD. We aim to investigate the striatal phospholipids (PLs) in hemiparkinsonism infused by rotenone in rats. As there are no cost-effective modes of PL, we have utilized dye-lipid complex technique for the first time in PD models for screening and also for semi-quantifying (individually) the levels of the deregulated PL in brain samples. Rats were divided into 2 groups: i. control and ii. ROT-infused which received intracranial injection of Rotenone (6 μg/μl; flow rate 0.2 μl/min). At the end of experimental period of 14 days, the striatum was dissected out for the analyses of PLs. Dye-based detection of PL and two-dimensional thin-layer chromatographic analyses of PL were performed. Detection of dye-PL complex was possible for phosphatidyl choline (PC), phosphatidyl inositol (PI), and spingomyelin (SM) (but not for phosphatidyl ethanolamine-PE) using dyes victoria blue B, toluidine blue and ammonium ferrothiocyanate, respectively. Two-dimensional analyses of phospholipids confirmed the dye-PL complex and depicted significant reduction (p < 0.05) on semi-quantitative assessment, in the striatum of control and hemiparkinsonic rats. We suggest a low level of PLs of PI in striatum of rats using a simple dye-detection that was validated by HR-LCMS. The finding implies that a critical role is being played by these PLs (PC, PI and SM) mainly PI (p < 0.001), in rotenone infused hemiparkinsonism, thus deserving wider but simpler investigations to detect and identify their role in parkinsonism.

摘要

帕金森病(PD)是一种进行性神经退行性疾病,其特征是黑质(SN)中多巴胺能神经元丧失以及运动协调能力丧失。主要脂质如磷脂的代谢受损,而磷脂在细胞功能和信号传导中起调节作用,这与PD的病理过程有关。我们旨在研究鱼藤酮注入大鼠后偏侧帕金森病模型纹状体中的磷脂(PLs)。由于没有经济有效的磷脂检测方法,我们首次在PD模型中使用染料 - 脂质复合物技术进行筛选,并对脑样本中失调的磷脂水平进行半定量(单独)分析。大鼠分为两组:i. 对照组;ii. 鱼藤酮注入组,接受颅内注射鱼藤酮(6μg/μl;流速0.2μl/min)。在14天的实验期结束时,取出纹状体进行磷脂分析。进行了基于染料的磷脂检测和磷脂的二维薄层色谱分析。分别使用维多利亚蓝B、甲苯胺蓝和硫氰酸铁铵染料,可检测到磷脂酰胆碱(PC)、磷脂酰肌醇(PI)和鞘磷脂(SM)(但不能检测磷脂酰乙醇胺 - PE)的染料 - 磷脂复合物。磷脂的二维分析证实了染料 - 磷脂复合物,并在半定量评估中显示,对照组和偏侧帕金森病大鼠纹状体中的磷脂水平显著降低(p < 0.05)。我们通过一种简单的染料检测方法表明,大鼠纹状体中PI的磷脂水平较低,该方法已通过高分辨率液相色谱 - 质谱联用(HR - LCMS)验证。这一发现表明,这些磷脂(PC、PI和SM),主要是PI(p < 0.001),在鱼藤酮注入的偏侧帕金森病中起着关键作用,因此值得进行更广泛但更简单的研究,以检测和确定它们在帕金森病中的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31a1/6084873/a4890579d517/gr1.jpg

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