Fasudil attenuates syncytin-1-mediated activation of microglia and impairments of motor neurons and motor function in mice

IF 3.5 4区医学 Q2 CHEMISTRY, MEDICINAL

Drug Development Research Pub Date : 2024-09-05 DOI:10.1002/ddr.22254

Mei Mao, Wen Zeng, Yan Zheng, Wen Fan, Yuanrong Yao

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Abstract

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease. Syncytin-1 (Syn), an envelope glycoprotein encoded by the env gene of the human endogenous retrovirus-W family, has been resorted to be highly expressed in biopsies from the muscles from ALS patients; however, the specific regulatory role of Syn during ALS progression remains uncovered.

In this study, C57BL/6 mice were injected with adeno-associated virus-overexpressing Syn, with or without Fasudil administration. The Syn expression was assessed by quantitative real-time polymerase chain reaction and immunohistochemistry analysis. The histological change of anterior tibial muscles was determined by hematoxylin-eosin staining. Qualitative ultrastructural analysis of electron micrographs obtained from lumbar spinal cords was carried out. Serum inflammatory cytokines were assessed by enzyme linked immunosorbent assay (ELISA) assay and motor function was recorded using Basso, Beattie, and Bresnahan (BBB) scoring, climbing test and treadmill running test. Immunofluorescence and western blot assays were conducted to examine microglial- and motor neurons-related proteins.

Syn overexpression significantly caused systemic inflammatory response, muscle tissue lesions, and motor dysfunction in mice. Meanwhile, Syn overexpression promoted the impairment of motor neuron, evidenced by the damaged structure of the neurons and reduced expression of microtubule-associated protein 2, HB9, neuronal nuclei and neuron-specific enolase in Syn-induced mice. In addition, Syn overexpression greatly promoted the expression of CD16/CD32 and inducible nitric oxide synthase (M1 phenotype markers), and reduced the expression of CD206 and arginase 1 (M2 phenotype markers). Importantly, the above changes caused by Syn overexpression were partly abolished by Fasudil administration.

This study provides evidence that Syn-activated microglia plays a pivotal role during the progression of ALS.

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法舒地尔可减轻 syncytin-1 介导的小胶质细胞激活以及小鼠运动神经元和运动功能损伤。

肌萎缩性脊髓侧索硬化症（ALS）是一种破坏性神经退行性疾病。Syncytin-1（Syn）是一种由人类内源性逆转录病毒-W家族 env 基因编码的包膜糖蛋白，在 ALS 患者的肌肉活检组织中高度表达。在这项研究中，C57BL/6小鼠注射了过表达Syn的腺相关病毒，同时服用或不服用法舒地尔。通过实时定量聚合酶链反应和免疫组化分析评估了Syn的表达。通过苏木精-伊红染色确定胫前肌的组织学变化。对腰脊髓电子显微镜照片进行了定性超微结构分析。血清炎症细胞因子通过酶联免疫吸附试验（ELISA）进行评估，运动功能通过巴索、比提和布雷斯纳汉（BBB）评分、攀爬试验和跑步机跑步试验进行记录。免疫荧光和 Western 印迹检测用于检查小胶质细胞和运动神经元相关蛋白。Syn过表达会明显导致小鼠全身炎症反应、肌肉组织损伤和运动功能障碍。同时，Syn过表达促进了运动神经元的损伤，表现为神经元结构受损，微管相关蛋白2、HB9、神经元核和神经元特异性烯醇化酶在Syn诱导的小鼠中表达减少。此外，Syn 的过表达大大促进了 CD16/CD32 和诱导型一氧化氮合酶（M1 表型标志物）的表达，降低了 CD206 和精氨酸酶 1（M2 表型标志物）的表达。重要的是，服用法舒地尔（Fasudil）可部分消除 Syn 过表达引起的上述变化。本研究提供的证据表明，Syn 激活的小胶质细胞在 ALS 的进展过程中起着关键作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Drug Development Research 医学-药学

CiteScore

6.40

自引率

2.60%

发文量

104

审稿时长

6-12 weeks

期刊介绍： Drug Development Research focuses on research topics related to the discovery and development of new therapeutic entities. The journal publishes original research articles on medicinal chemistry, pharmacology, biotechnology and biopharmaceuticals, toxicology, and drug delivery, formulation, and pharmacokinetics. The journal welcomes manuscripts on new compounds and technologies in all areas focused on human therapeutics, as well as global management, health care policy, and regulatory issues involving the drug discovery and development process. In addition to full-length articles, Drug Development Research publishes Brief Reports on important and timely new research findings, as well as in-depth review articles. The journal also features periodic special thematic issues devoted to specific compound classes, new technologies, and broad aspects of drug discovery and development.