Institute of Integrative Medicine, Department of Integrated Traditional Chinese and Western Medicine, Xiangya Hospital, Central South University, 410008, Changsha, PR China.
Xiangya School of Medicine, Central South University, 410013, Changsha, China.
J Ethnopharmacol. 2020 Aug 10;258:112826. doi: 10.1016/j.jep.2020.112826. Epub 2020 Apr 13.
Xuefu Zhuyu decoction (XFZYD) is a traditional Chinese herbal prescription. It is effective in treating traumatic brain injury (TBI). However, the underlying molecular mechanisms remain unclear.
This study aimed to reveal the possible mechanisms of XFZYD in treating acute TBI through proteomics clues.
Controlled Cortical Impact (CCI) rats were given gavage administration of XFZYD (9 g/kg/d) or distilled water (equal volume) for three days. The Modified Neurological Severity Score (mNSS), brain water content, HE staining, Nissl staining and immunohistochemistry were performed to assess the effects of XFZYD for TBI treatment. Additionally, tandem mass tag-based (TMT) quantitative proteomics technology was applied to detect proteins of brain cortex. Bioinformatics analysis including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and Protein-protein interaction (PPI) networks were used to analyze differentially expressed proteins (DEPs). Bioinformatics Analysis Tool for Molecular mechanism of TCM (BATMAN-TCM) was conducted to anchor diseases and pathways. Besides, western blotting and immunofluorescence were exerted to verify related proteins.
XFZYD improved neurologic functions, reduced encephaledema and ameliorated cell morphology around the injured area in CCI rats. A total of 6099 proteins were identified with false discovery rate (FDR) < 1%. Overlapping DEPs (105 DEPs) were identified (295 DEPs and 804 DEPs in CCI/Sham or XFZYD/CCI group, respectively). Of these DEPs, 17 were regulated by XFZYD. Bioinformatics analysis showed that the 17 DEPs were predominantly related to platelet activation and PI3K-Akt signaling pathway. Next, PLG and CD34 were verified with molecular biotechnology.
XFZYD exerts therapeutic effects through multi-pathways regulation in the treatment of TBI. This work may provide proteomics clues for the continuation of research on TBI treatment with XFZYD.
血府逐瘀汤（XFZYD）是一种传统的中药方剂，对治疗创伤性脑损伤（TBI）有显著疗效。但其潜在的分子机制尚不清楚。
本研究旨在通过蛋白质组学线索揭示 XFZYD 治疗急性 TBI 的可能机制。
采用控制性皮质撞击（CCI）大鼠模型，给予 XFZYD（9g/kg/d）或蒸馏水（等体积）灌胃 3 天。采用改良神经功能缺损评分（mNSS）、脑水含量、HE 染色、尼氏染色和免疫组织化学等方法评估 XFZYD 对 TBI 的治疗作用。此外，采用串联质量标签（TMT）定量蛋白质组学技术检测皮质蛋白质。利用基因本体论（GO）、京都基因与基因组百科全书（KEGG）通路和蛋白质-蛋白质相互作用（PPI）网络等生物信息学分析方法对差异表达蛋白（DEPs）进行分析。采用中药分子机制生物信息分析工具（BATMAN-TCM）对疾病和通路进行锚定。此外，还进行了 Western blot 和免疫荧光实验来验证相关蛋白。
XFZYD 可改善 CCI 大鼠的神经功能，减轻脑水肿，改善损伤区周围细胞形态。假发现率（FDR）<1%时共鉴定出 6099 种蛋白质。重叠的 DEPs（CCI/Sham 或 XFZYD/CCI 组分别有 105 个和 804 个 DEPs）。其中，17 个受 XFZYD 调控。生物信息学分析表明，这 17 个 DEPs 主要与血小板激活和 PI3K-Akt 信号通路相关。接下来，通过分子生物技术验证了 PLG 和 CD34。
XFZYD 通过多途径调节在 TBI 治疗中发挥治疗作用。这项工作可能为 XFZYD 治疗 TBI 的研究提供蛋白质组学线索。
