银杏二萜内酯葡胺注射液对缺血性脑卒中小鼠黑质脑区的调控机制研究  

Regulatory mechanism of Diterpene Ginkgolides Meglumine Injection on substantia nigra brain region in ischemic stroke mice

作　　者：刘子宇 徐欣玉 吕文欣 郭志宏 张新庄[2] 曹亮[2] 王振中[1,2] 田金洲 王团结[2] 潘贤 武子寅 肖伟[1,2] LIU Ziyu;XU Xinyu;LYU Wenxin;GUO Zhihong;ZHANG Xinzhuang;CAO Liang;WANG Zhenzhong;TIAN Jinzhou;WANG Tuanjie;PAN Xian;WU Ziyin;XIAO Wei(State Key Laboratory on Technologies for Chinese Medicine Pharmaceutical Process Control and Intelligent Manufacture,Nanjing University of Traditional Chinese Medicine,Nanjing 210023,China;Jiangsu Kanion Pharmaceutical Co.,Ltd.,Lianyungang 222001,China)

机构地区：[1]南京中医药大学,中药制药过程控制与智能制造技术全国重点实验室,江苏南京210023 [2]江苏康缘药业股份有限公司,江苏连云港222001

出　　处：《中草药》2024年第11期3735-3748,共14页Chinese Traditional and Herbal Drugs

基　　金：江苏省院士工作站(BM2023118);连云港市2023年度第六期“521工程”科研资助项目(LYG06521202302);连云港市自然科学基金资助项目(JCYJ2323);江苏省自然科学青年基金资助项目(BK20210139)。

摘　　要：目的研究银杏二萜内酯葡胺注射液(Diterpene Ginkgolides Meglumine Injection,DGMI)在C57BL/6J小鼠大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)模型黑质脑区中抗脑缺血再灌注损伤的潜在作用通路。方法C57BL/6J小鼠按随机数字表分为假手术组、模型组、DGMI(25 mg/kg)组及银杏叶提取物761(Ginkgo biloba extract-761,EGb-761,100 mg/kg)组,构建MCAO模型,通过测定各组小鼠术后24 h的改良版神经功能缺损评分和脑梗死率,评价DGMI相较于EGb-761治疗缺血性脑卒中的疗效。进一步利用Illumina二代测序平台对各组小鼠脑组织黑质样本进行高通量转录组测序,并结合基因集富集分析(gene set enrichment analysis,GSEA)、差异表达基因(differentially expressed genes,DEGs)的基因本体(gene ontology,GO)功能和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析等生物信息学分析方法,评价DGMI抗脑缺血再灌注损伤作用及潜在作用机制。采用qRT-PCR验证转录组测序中关键基因表达。结果与假手术组比较,模型组改良版神经功能缺损评分和脑梗死率显著升高(P<0.001);与模型组比较,DGMI组治疗效果与EGb-761接近,可显著改善改良版神经功能缺损评分和脑梗死率(P<0.01、0.001)。转录组的数据降维分析显示,模型组与假手术组样本显著分离。GSEA结果显示,模型组与假手术组对比显著富集35条通路,DGMI组相较于模型组可有效逆转7条通路。与假手术组相比,模型组共筛选出88个DEGs,将其与他人发现的大小鼠MCAO模型组共同差异基因比较(15个),有10个基因相同。模型组GO功能分析主要富集在炎症和细胞趋化等过程,KEGG通路分析主要富集在细胞因子互作、肿瘤坏死因子等信号通路。与模型组相比,DGMI组DEGs的GO功能分析主要富集在神经信号传导过程,KEGG通路分析主要富集在多巴胺能信号通路、神经活性受体配体作用等。qRT-PCR�Objective To study the effect of Diterpene Ginkgolides Meglumine Injection(DGMI)on middle cerebral artery occlusion(MCAO)in C57BL/6J mice and investigate the potential pathways of anti-cerebral ischemia-reperfusion injury in substantial nigra(SN)brain region of MCAO model.Methods C57BL/6J mice were randomly divided into sham group,model group,DGMI(25 mg/kg)group and Ginkgo biloba extract-761(EGb-761,100 mg/kg)group,MCAO model was established.The efficacy of DGMI compared with EGb-761 in the treatment of ischemic stroke was evaluated by measuring modified neurological severity score and cerebral infarction rate at 24 h after MCAO.The Illumina next-generation sequencing platform was further used to perform high-throughput transcriptome sequencing of substantia nigra samples of the brain tissue of each group.The effect and potential mechanism of DGMI against cerebral ischemia-reperfusion injury was explored by bioinformatics analysis methods such as gene set enrichment analysis(GSEA),gene ontology(GO)function and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis of differentially expressed genes(DEGs).qRT-PCR was used to verify the expressions of key genes in transcriptome sequencing.Results Compared with sham group,modified neurological severity score and cerebral infarction rate in model group were significantly increased(P<0.001);Compared with model group,the treatment effect of DGMI group was similar to that of EGb-761,which significantly improved the modified neurological severity score and cerebral infarction rate(P<0.01,0.001).Data dimensionality reduction analysis of transcriptome showed that model group was significantly separated from control group.GSEA results showed that 35 pathways were significantly enriched in model group compared with sham group,and seven pathways were effectively reversed in DGMI group compared with model group.Compared with sham group,a total of 88 DEGs were screened out in model group.There were 10 same genes compared with 15 common DEGs in rat and mouse MCA

关 键 词：银杏二萜内酯葡胺注射液 缺血性脑卒中 黑质 大脑中动脉闭塞模型 转录组测序 

分 类 号：R285.5[医药卫生—中药学]