摘要
目的:在确定谷红注射液保护脑缺血的基础上,采用RNA-Seq转录组测序与生物信息分析的方法探究谷红注射液抗脑缺血的分子机制。方法:采用线栓法制备大脑中动脉栓塞(MCAO)模型,设置假手术组,模型组,谷红注射液低、中剂量组(0.625,2.5 m L·kg^-1·d^-1),阳性药组(金纳多注射液,8 m L·kg^-1·d^-1),通过Ludmila Belayev 12分评分法进行药效学评价,并采用RNA-Seq技术检测药物干预前后的差异基因,使用DAVID,String及The Human Phenotype Ontology等数据库,进行富集分析、聚类分析、以及与脑缺血疾病靶标的关联分析,通过Cytoscape3.4.0构建相关的调控网络。结果:与假手术组相比,模型组大鼠可见明显神经功能缺损(P<0.01);与模型组比较,谷红注射液高剂量组减轻了大鼠神经功能损伤(P<0.05)。转录组数据分析表明谷红注射液主要是通过调控细胞凋亡,炎症反应,氧化应激,Toll样受体信号通路及丝裂原激活的蛋白激酶(MAPK)信号通路等生物学过程干预脑缺血。此外,谷红液射液干预脑缺血的差异基因关联到20个脑缺血疾病相关的靶标,且关联到64个MAPK信号通路相关的基因,其中23个基因参与凋亡与炎症等过程。结论:谷红注射液通过多条途径发挥抗脑缺血作用,其中MAPK信号通路是其发挥抗凋亡及炎症作用的重要途径。
Objective:Based on the protective effect of Guhong injection(GH)on cerebral ischemia,mechanism of GH against cerebral ischemia was identified using RNA-seq transcriptome and bioinformation analysis.Method:The model of middle cerebral artery occlusion(MCAO)was established through thread embolization.Sham group,model group,low-dose GH group(0.625 m L·kg^-1·d^-1),high-dose GH group(2.5 m L·kg^-1·d^-1),positive group(Ginaton,8 m L·kg-1·d-1)were set up.Ludmila Belayev 12-point scoring method was applied to assess the protective effect of GH against MCAO-induced cerebral ischemia.And the differentially expressed genes after treatment with GH were identified by RNA-Seq technology.Enrichment analysis,cluster analysis and association analysis on disease targets of cerebral ischemia were carried out through such databases as DAVID,String and The Human Phenotype Ontology.Finally,the regulatory network was constructed by Cytoscape3.4.0.Result:Compared with the sham group,the neurological impairment was obvious in the model group(P<0.01),and the neurological impairment was alleviated in the GH group compared with the model group(P<0.05).RNA-Seq technology analysis showed that GH regulated genes involving such biological processes as cell apoptosis,inflammation,oxidative stress,toll-like signaling pathway and mitogen-activated protein kinase(MAPK)signaling pathway.Twenty disease targets and 64 MAPK signaling pathway genes were associated with differentially expressed genes after GH treatment,in which 23 genes were involved in apoptosis and inflammation.Conclusion:GH protected against cerebral ischemia in many ways,among which MAPK signaling pathway is an important way to exert its effect in inhibiting apoptosis and inflammation.
作者
周瑞
项昌培
高金环
张毅
张晶晶
杨洪军
ZHOU Rui;XIANG Chang-pei;GAO Jin-huan;ZHANG Yi;ZHANG Jing-jing;YANG Hong-jun(School of Traditional Chinese Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第7期195-203,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81974550,81330086,81603422).
作者简介
第一作者:周瑞,在读硕士,从事中药药理与应用研究,E-mail:zhourui677000@163.com;通信作者:张晶晶,博士,副研究员,从事中药心脑血管分子药理学研究,E-mail:zjj4785@163.com;通信作者:杨洪军,博士,研究员,从事中药心脑血管药理研究及中药大品种培育研究,Tel:010-64032656,E-mail:hongjun0420@vip.sina.com
