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东莨菪碱诱导的失忆小鼠模型海马中差异微小RNA表达的分析

Analysis of Differential microRNA Expression in the Hippocampus of Scopolamine-Induced Amnesic Mouse Model.

作者信息

Verma Samita, Sivanandam Thamil Mani

机构信息

Biochemistry and Molecular Biology Laboratory, Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India.

出版信息

Mol Neurobiol. 2025 Apr;62(4):4881-4898. doi: 10.1007/s12035-024-04573-y. Epub 2024 Nov 4.

Abstract

Amnesia is characterized by memory deficits linked to various neurodegenerative pathologies and can be induced by the administration of scopolamine, a cholinergic antagonist. Scopolamine-induced amnesia is a well-studied pharmacological animal model that simulates memory impairment caused by aging, brain illnesses, neuropathologies, and trauma. However, the molecular mechanism of amnesia, more importantly in terms of microRNA (miRNA) regulation, is not well understood. Therefore, this study aimed to analyze miRNA profiles in the hippocampus of both control mice and those treated with scopolamine (amnesic mice). Initially, a short cDNA library was prepared for each sample and then sequenced on the Illumina platform. Among the total differentially expressed miRNAs, 113 were significantly upregulated and 96 were downregulated in the scopolamine group in comparison to the control group. Ten upregulated and ten downregulated miRNAs were validated to confirm the reliability of the sequencing results using qRT-PCR (quantitative real-time PCR). Furthermore, we performed a target prediction analysis intersecting the results from TargetScan, miRDB (miRNA database), and Miranda to analyze the targets of the dysregulated miRNAs. We also conducted a pathway analysis to investigate the molecular, cellular, and biological functions of these targets. miRNA‒target interactions were found to play roles in various signaling pathways during amnesia. These results provide an initial insight for the contribution of miRNAs to scopolamine-induced amnesia, as well as their possible application as markers of disease pathology.

摘要

失忆症的特征是与各种神经退行性病变相关的记忆缺陷,并且可由胆碱能拮抗剂东莨菪碱诱导产生。东莨菪碱诱导的失忆症是一种经过充分研究的药理学动物模型,可模拟由衰老、脑部疾病、神经病理学和创伤引起的记忆障碍。然而,失忆症的分子机制,尤其是在 microRNA(miRNA)调控方面,尚未得到充分理解。因此,本研究旨在分析对照小鼠和接受东莨菪碱治疗的小鼠(失忆小鼠)海马体中的 miRNA 谱。最初,为每个样本制备了一个短 cDNA 文库,然后在 Illumina 平台上进行测序。在总共差异表达的 miRNAs 中,与对照组相比,东莨菪碱组中有 113 个显著上调,96 个下调。使用 qRT-PCR(定量实时 PCR)验证了 10 个上调和 10 个下调的 miRNAs,以确认测序结果的可靠性。此外,我们进行了靶标预测分析,将 TargetScan、miRDB(miRNA 数据库)和 Miranda 的结果进行交叉分析,以分析失调 miRNAs 的靶标。我们还进行了通路分析,以研究这些靶标的分子、细胞和生物学功能。发现 miRNA-靶标相互作用在失忆症期间的各种信号通路中发挥作用。这些结果为 miRNAs 对东莨菪碱诱导的失忆症的贡献及其作为疾病病理学标志物的可能应用提供了初步见解。

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