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振宝丸通过调节 GPR17 的表达来防治急性脊髓损伤。

Zhenbao pill protects against acute spinal cord injury via regulating the expression of GPR17.

机构信息

Department of Spine Surgery, Inner Mongolia People's Hospital, Hohhot 010017, Inner Mongolia, People's Republic of China.

Department of Clinic, Inner Mongolia Medical University People's Hospital, Hohhot 010020, Inner Mongolia, People's Republic of China.

出版信息

Biosci Rep. 2018 Jan 19;38(1). doi: 10.1042/BSR20171132. Print 2018 Feb 28.

Abstract

The aim of the present study was to observe the effect of zhenbao pill on the motor function of acute spinal cord injury (ASCI) rats and the molecular mechanisms involving and G-protein-coupled receptor 17 (GPR17). ASCI rat model was established by modified Allen method, and then the rats were divided into three groups. SH-SY5Y cells were cultured overnight in hypoxia condition and transfected with mimic or inhibitor. The hind limb motor function of the rats was evaluated by Basso, Beattie, Bresnahan (BBB) scoring system. Quantitative real-time PCR (qRT-PCR) and Western blot were used to detect the expression of , GPR17, inducible nitric oxide synthase (iNOS), interleukin 1β (IL-1β), and tumor necrosis factor α (TNF-α). Neuronal apoptosis was measured using flow cytometry assay. Luciferase reporter assay was performed to determine the regulation of on GPR17. Zhenbao pill could enhance hind limb motor function and attenuate the inflammatory response caused by ASCI. Moreover, zhenbao pill increased the level of and decreased GPR17 expression Bioinformatics software predicted that GPR17 3'-UTR had a binding site with Luciferase reporter assay showed that had a negative regulatory effect on GPR17 expression. Knockdown of could reverse the effect of zhenbao pill on the up-regulation of GPR17 induced by hypoxia, reversed the inhibitory effect of zhenbao pill on the cell apoptosis induced by hypoxia and the recovery of zhenbao pill on hind limb motor function in ASCI rats. Zhenbao pill could inhibit neuronal apoptosis by regulating /GPR17 expression, and then promoting the recovery of spinal cord function.

摘要

本研究旨在观察振脑丸对急性脊髓损伤(ASCI)大鼠运动功能的影响及其涉及的分子机制和 G 蛋白偶联受体 17(GPR17)。采用改良 Allen 法建立 ASCI 大鼠模型,将大鼠分为三组。SH-SY5Y 细胞在缺氧条件下培养过夜,并转染 模拟物或抑制剂。采用 Basso、Beattie、Bresnahan(BBB)评分系统评估大鼠后肢运动功能。采用实时定量 PCR(qRT-PCR)和 Western blot 检测大鼠 、GPR17、诱导型一氧化氮合酶(iNOS)、白细胞介素 1β(IL-1β)和肿瘤坏死因子 α(TNF-α)的表达。采用流式细胞术检测神经元凋亡。采用荧光素酶报告基因检测 对 GPR17 的调控作用。振脑丸可增强后肢运动功能,减轻 ASCI 引起的炎症反应。此外,振脑丸增加了 的水平,并降低了 GPR17 的表达。生物信息学软件预测 GPR17 3'-UTR 与 有结合位点。荧光素酶报告基因检测显示,对 GPR17 表达具有负调控作用。下调 可逆转振脑丸对缺氧诱导的 GPR17 上调的作用,逆转振脑丸对缺氧诱导的细胞凋亡的抑制作用和振脑丸对 ASCI 大鼠后肢运动功能的恢复作用。振脑丸通过调节 /GPR17 表达抑制神经元凋亡,进而促进脊髓功能恢复。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66bf/5773823/d6e27d66ce5a/bsr-38-bsr20171132-g1.jpg

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