School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China.
School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 100029, China.
J Ethnopharmacol. 2020 Mar 1;249:112362. doi: 10.1016/j.jep.2019.112362. Epub 2019 Oct 29.
There is a lack of systematic descriptions and characterization of strokes and their effects in both the cerebral hippocampus and cortex. Shuxuetong (SXT) injection was reported to have good therapeutic effects in the clinic; therefore, it was selected as a drug intervention method for cerebral ischemia repair in rat models. The aim of this study was to understand the features of molecules and pathways and to reveal key processes of SXT repair.
Evaluation of neurological deficit and infarct volume measurement was used to estimate the pharmacological effects of SXT injection on Ischemia-reperfusion(I/R) model rats. LC-MS/MS and RNA-Seq analysis were used to analyze the proteins and mRNA expression in the cerebral hippocampus and cortex 6 h and 24 h after ischemic injury and repair. A label-free approach (IBAQ) for proteomics analysis and FPKM based on gene read count for transcriptomics analysis were used to quantify the differences among the three experimental groups (Sham, Model and SXT-treated groups). Transcriptomics and proteomics analyses were verified by RT-qPCR and western blotting.
By combining LC-MS/MS and RNA-Seq, eight larger datasets (two time points and two tissues) were confidently identified in more than three biological replicates. An average of 4500 unique proteins and 8200 protein-coding genes were confidently identified. By combining the subcellular localization, hierarchical clustering, pathway enrichment analysis in the injury and repair phase, six core proteins and related genes that were significantly expressed were verified as candidates for cerebral ischemic injury by western blotting and quantitative real-time PCR. Meanwhile, the results indicated that there was better expression in the 6 h group by significant proteomics analysis during the development and progression of cerebral ischemia. Two primary co-enriched pathways, the PI3K-AKT and MAPK signaling pathways, and six related core candidates may play key roles in molecular mechanisms related to cerebral ischemic injury and repair by SXT injection.
Our data not only identified six core candidates and two key signaling pathways for cerebral ischemic injury and verification but also provided evidence for the explanation, prevention and treatment of cerebral ischemia by SXT injection. The results of the present study provide evidence for the explanation, prevention and treatment of cerebral ischemia by SXT injection.
目前缺乏对脑卒中及其在大脑海马体和皮质中的影响的系统描述和特征分析。疏血通注射液（SXT）在临床上有良好的治疗效果，因此被选择作为大鼠脑缺血修复的药物干预方法。本研究旨在了解分子和途径的特征，并揭示 SXT 修复的关键过程。
采用神经功能缺损评分和脑梗死体积测量评估 SXT 注射液对缺血再灌注（I/R）模型大鼠的药效。采用 LC-MS/MS 和 RNA-Seq 分析方法，分别对缺血损伤后 6 h 和 24 h 大脑海马体和皮质的蛋白质和 mRNA 表达进行分析。采用无标记定量（IBAQ）方法和基于基因读段的 FPKM 方法对三组实验（假手术组、模型组和 SXT 处理组）进行蛋白质组学和转录组学分析。通过 RT-qPCR 和 Western blot 验证转录组学和蛋白质组学分析结果。
通过结合 LC-MS/MS 和 RNA-Seq，在超过三个生物学重复中，共鉴定了八个较大的数据集（两个时间点和两个组织）。平均鉴定到 4500 个独特的蛋白质和 8200 个蛋白质编码基因。通过结合亚细胞定位、损伤和修复阶段的层次聚类、通路富集分析，通过 Western blot 和定量实时 PCR 验证了 6 个核心蛋白和相关基因作为脑缺血损伤的候选物。同时，通过显著的蛋白质组学分析，在脑缺血发生发展过程中，结果表明 6 h 组表达更好。PI3K-AKT 和 MAPK 信号通路两个主要的共富集通路，以及 6 个相关的核心候选物，可能通过 SXT 注射在与脑缺血损伤和修复相关的分子机制中发挥关键作用。
本研究不仅鉴定了 6 个核心候选物和 2 个关键信号通路在脑缺血损伤和验证中的作用，还为疏血通注射液治疗脑缺血提供了证据。本研究结果为疏血通注射液治疗脑缺血提供了解释、预防和治疗的证据。
