Zhou Xiu-Min, Liu Jie, Wang Ying, Zhang Shu-Li, Zhao Xin, Xu Xiang, Pei Jian, Zhang Man-He
Department of Anesthesiology, Tangshan Gongren Hospital, Tangshan, China.
Department of Neurosurgery, Tangshan Gongren Hospital, Tangshan, China.
J Cell Biochem. 2019 May;120(5):6908-6919. doi: 10.1002/jcb.26704. Epub 2018 Jan 27.
Our study aims to elucidate the mechanisms how microRNA-129-5p (miR-129-5p) involved in the neuroprotective effect of dexmedetomidine (DEX) on hypoxic-ischemic brain injury (HIBI) by targeting the type III procollagen gene (COL3A1) through the Wnt/β-catenin signaling pathway in neonatal rats. A total of 120 rats were obtained, among which 15 rats were selected as sham group and rest rats as model, DEX, DEX + negative control (DEX + NC), DEX + miR-129-5p mimics, DEX + miR-129-5p inhibitors, DEX + XAV-939, and DEX + miR-129-5p inhibitors + XAV-939 groups. A dual-luciferase reporter assay was performed for the target relationship between miR-129-5p and COL3A1. Weight rate and water content of cerebral hemisphere were detected. Quantitative real-time polymerase chain reaction and Western blot analysis were conducted to detect miR-129-5p expression and expressions of COL3A1, E-cadherin, T-cell factor (TCF)- 4, and β-catenin. The DEX, DEX + miR-129-5p mimics, DEX + XAV-939 groups had increased weight rate of the cerebral hemisphere, but decreased water content of left cerebral hemisphere, levels of COL3A1, β-catenin, TCF-4, and E-cadherin in the hippocampus compared with the model and DEX + miR-129-5p inhibitors groups. COL3A1 was verified as the target gene of the miR-129-5p. Compared with the DEX + NC and DEX + miR-129-5p inhibitors + XAV-939 groups, the DEX + XAV-939 and DEX + miR-129-5p mimics groups had elevated weight rate of the cerebral hemisphere, but reduced water content of left cerebral hemisphere, levels of COL3A1, β-catenin, TCF-4, and E-cadherin in the hippocampus. Our findings demonstrate that miR-129-5p improves the neuroprotective role of DEX in HIBI by targeting COL3A1 through the Wnt/β-catenin signaling pathway in neonatal rats.
我们的研究旨在阐明微小RNA-129-5p(miR-129-5p)如何通过Wnt/β-连环蛋白信号通路靶向III型前胶原基因(COL3A1),参与右美托咪定(DEX)对新生大鼠缺氧缺血性脑损伤(HIBI)的神经保护作用。共获得120只大鼠,其中15只作为假手术组,其余作为模型组、DEX组、DEX+阴性对照(DEX+NC)组、DEX+miR-129-5p模拟物组、DEX+miR-129-5p抑制剂组、DEX+XAV-939组以及DEX+miR-129-5p抑制剂+XAV-939组。对miR-129-5p与COL3A1之间的靶向关系进行双荧光素酶报告基因检测。检测脑半球的重量率和含水量。采用定量实时聚合酶链反应和蛋白质免疫印迹分析检测miR-129-5p表达以及COL3A1、E-钙黏蛋白、T细胞因子(TCF)-4和β-连环蛋白的表达。与模型组和DEX+miR-129-5p抑制剂组相比,DEX组、DEX+miR-129-5p模拟物组、DEX+XAV-939组脑半球重量率增加,但左脑半球含水量、海马中COL3A1、β-连环蛋白、TCF-4和E-钙黏蛋白水平降低。COL3A1被证实为miR-129-5p的靶基因。与DEX+NC组和DEX+miR-129-5p抑制剂+XAV-939组相比,DEX+XAV-939组和DEX+miR-129-5p模拟物组脑半球重量率升高,但左脑半球含水量、海马中COL3A1、β-连环蛋白、TCF-4和E-钙黏蛋白水平降低。我们的研究结果表明,miR-129-5p通过在新生大鼠中通过Wnt/β-连环蛋白信号通路靶向COL3A1,改善了DEX对HIBI的神经保护作用。