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不宁腿综合征动物模型中的蛋白酪氨酸磷酸酶受体δ(PTPRD)基因

Protein tyrosine phosphatase receptor type delta (PTPRD) gene in an animal model of restless legs syndrome.

作者信息

Morais Milca Abda, Franco Beatriz Silva, Holanda Alessandro Spencer Souza, de Paula Simino Laís Angélica, Veras Alana Carolina Costa, Torsoni Marcio Alberto, Manconi Mauro, Torsoni Adriana Souza, Esteves Andrea Maculano

机构信息

Faculdade de Ciências Aplicadas, Universidade Estadual de Campinas, Limeira, Brazil.

Faculdade de Educação Física, Universidade Estadual de Campinas, Campinas, Brazil.

出版信息

J Sleep Res. 2023 Apr;32(2):e13716. doi: 10.1111/jsr.13716. Epub 2022 Sep 2.

Abstract

The pathophysiology of the restless legs syndrome (RLS) is related to dopaminergic dysfunction, reduced iron and variations in gene expression, such as the protein tyrosine phosphatase receptor type delta gene (PTPRD). Animal models could be key to achieving a mechanistic understanding of RLS and to facilitate efficient platforms for evaluating new therapeutics. Thus, the aim of this study was to evaluate the expression of PTPRD, of genes and proteins associated with RLS, the sleep patterns and the cardiovascular parameters in an animal model of RLS (spontaneously hypertensive rat [SHR]). Rats were divided into two groups: (i) Wistar-Kyoto and (ii) SHR. Cardiovascular parameters were assessed by tail plethysmography. Polysomnography was used to analyse the sleep pattern (24 h). For the PTPRD analyses, quantitative polymerase chain reaction (qPCR) and indirect enzyme-linked immunosorbent assay (ELISA) techniques were used. To evaluate the tyrosine hydroxylase enzyme, dopamine transporter (DAT) and type 2 dopaminergic receptor, qPCR and Western Blotting techniques were used. For the quantification of iron, ferritin and transferrin, the ELISA method was used. SHRs had higher blood pressure, alterations in sleep pattern, lower expression of protein content of PTPRD, lower expression of DAT, and lower serum concentrations of ferritin. These data suggest that the behavioural, physiological, and molecular changes observed in SHRs provide a useful animal model of RLS, reinforcing the importance of this strain as an animal model of this sleep disorder.

摘要

不宁腿综合征(RLS)的病理生理学与多巴胺能功能障碍、铁缺乏及基因表达变化有关,如δ型蛋白酪氨酸磷酸酶受体基因(PTPRD)。动物模型可能是实现对RLS进行机制性理解并促进评估新疗法的有效平台的关键。因此,本研究的目的是评估RLS动物模型(自发性高血压大鼠[SHR])中PTPRD的表达、与RLS相关的基因和蛋白质表达、睡眠模式及心血管参数。大鼠分为两组:(i)Wistar-Kyoto大鼠和(ii)SHR大鼠。通过尾容积描记法评估心血管参数。采用多导睡眠图分析睡眠模式(24小时)。对于PTPRD分析,使用定量聚合酶链反应(qPCR)和间接酶联免疫吸附测定(ELISA)技术。为评估酪氨酸羟化酶、多巴胺转运体(DAT)和2型多巴胺能受体,使用qPCR和蛋白质印迹技术。采用ELISA法对铁、铁蛋白和转铁蛋白进行定量。SHR大鼠血压更高、睡眠模式改变、PTPRD蛋白含量表达降低、DAT表达降低以及血清铁蛋白浓度降低。这些数据表明,在SHR大鼠中观察到的行为、生理和分子变化提供了一种有用的RLS动物模型,强化了该品系作为这种睡眠障碍动物模型的重要性。

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