网络药理学联合circRNA研究凉血通瘀方干预脑出血急性期药效机制  

Network Pharmacology Combined with circRNA to Study the Pharmacodynamic Mechanism of Liangxue Tongyu Prescription in Intervening in the Acute Phase of Intracerebral Hemorrhage

作　　者：李薇懿 王燕[1] 李国春[2] LI Weiyi;WANG Yan;LI Guochun(The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;Department of Public Health,Nanjing University of Traditional Chinese Medicine,Nanjing 210023,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450000 [2]南京中医药大学公共卫生系,南京210023

出　　处：《世界科学技术-中医药现代化》2024年第9期2254-2264,共11页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会面上项目(81373512):急性脑出血瘀热病机单元的动态测量及其生物标志物的研究,负责人:李国春。

摘　　要：目的通过研究凉血通瘀方对于自发性高血压大鼠(Spontaneously hypertensive rats,SHR)脑出血急性期环状RNA(circRNA)的作用靶点及可能相关信号通路,探讨凉血通瘀方治疗脑出血急性期的药效机制。方法将SHR分成模型组、实验组及空白组,模型组、实验组构建脑出血SHR模型,并对SHR神经功能损伤进行评分。术后实验组给予凉血通瘀方,模型组、空白组给予等剂量生理盐水,各5天。借助高通量测序技术检测3组SHR脑组织样本中circRNA,对显著差异表达的circRNA进行聚类分析、GO和KEGG富集分析。使用实时荧光定量聚合酶链式反应(Real-time PCR,RT-PCR)验证差异倍数排名前5的显著差异circRNA的表达情况。将显著差异表达的circRNA转化为人类同源基因,结合课题组前期筛选的脑出血急性期治疗靶点,得到交集靶点输入Cytoscape软件构建中药-成分-靶点-通路网络。结果测序结果为显著差异表达的circRNA共有150个,其中60个为上调、90个为下调circRNA。RT-PCR验证结果显示前5名circRNA的表达趋势与高通量测序结果一致。网络药理学结果显示,凉血通瘀方中赤芍、大黄、地黄、牡丹皮、石菖蒲和三七6味中药里多种活性成分对应脑出血急性期治疗靶点PRKCA,即CIRI_circ_10530的亲本基因。849条GO富集条目中有CIRI_circ_10530亲本基因分布的共209条;149条KEGG信号通路中CIRI_circ_10530亲本基因分布在62条信号通路中。其中大黄是凉血通瘀方中治疗脑出血急性期的重要中药,山柰酚、大黄素和槲皮素为药效机制的重要活性成分。结论凉血通瘀方干预脑出血急性期的作用机制可能与山柰酚、槲皮素和大黄素等关键活性成分激活PRKCA,并调控PKC影响其关键信号通路有关。Objective To explore the therapeutic mechanism of Liangxue Tongyu Prescription in the treatment of acute Intracerebral hemorrhage by studying the target of Liangxue Tongyu Prescription on circRNAand possible related signaling pathways in Spontaneously hypertensive rats(SHR)in the acute stage of Intracerebral hemorrhage.Methods The SHR was divided into model group,experimental group and blank group.Intracerebral hemorrhage models were performed in model group and experimental group,the neurological deficits of SHR were scored.The experimental group was given Liangxue Tongyu Prescription for 5 days,the model group and the blank group were given equal doses of normal saline for 5 days each.The circRNA in the three groups of SHR brain tissue samples were detected by highthroughput sequencing technology,and the significantly different circRNAs were analyzed by Cluster Analysis,Go and KEGG enrichment analysis.Real-time fluorescence quantitative polymerase chain reaction(RT-PCR)was used to verify the expressions of the top 5 significantly different circRNAwith fold change.The significantly differentially expressed circRNA were transformed into human homologous genes,and combined with the treatment targets selected by our research group in the early stage of Intracerebral hemorrhage,the intersection targets were input into Cytoscape software to construct the traditional Chinese medicines-components-targets-pathways network.Results The results of sequencing showed that there were 150 significantly different circRNA,including 60 up-regulated circRNAand 90 down-regulated circRNA.RT-PCR verification results showed that the expression trends of the top 5 circRNA were consistent with the high-throughput sequencing results.Network pharmacology results show that multiple active ingredients in six traditional Chinese medicines including Radix Paeoniae Rubra,Rheum officinale,Rehmannia glutinosa,Paeonia suffruticosa,Acorus tatarinowii and Panax notoginseng in Liangxue Tongyu Prescription correspond to the acute phase treatment targ

关 键 词：凉血通瘀方 脑出血急性期 circRNA 网络药理学 转录组学 

分 类 号：R-058[医药卫生]