摘要
目的采用RNA测序技术,从基因与分子层面探讨益肾养髓方治疗脊髓型颈椎病的潜在作用机制。方法30只SD大鼠随机分为假手术组、模型组和中药组,采用脊髓压迫法建立脊髓型颈椎病大鼠模型,中药组于术后1周开始予益肾养髓方灌胃,每日1次,连续8周。采用Basso Beattie Bresnahan(BBB)评分评估大鼠脊髓功能,透射电镜观察脊髓组织病理形态,RNA测序技术筛选各组差异基因并进行生物信息学分析,RT-PCR检测差异基因mRNA表达,Western blot检测差异基因蛋白表达。结果与假手术组比较,模型组大鼠BBB评分显著降低(P<0.05);与模型组比较,术后4周开始,中药组大鼠BBB评分显著升高(P<0.05)。透射电镜观察显示,模型组脊髓组织髓鞘不完整,可见断裂或卷曲,神经元线粒体嵴断裂、缺失;中药组髓鞘结构排列较规则,线粒体形态破坏较轻。RNA测序技术筛选出假手术组与模型组差异基因303个(153个上调、150个下调),模型组与中药组差异基因341个(130个上调、211个下调),对差异表达基因取交集,得到117个共表达基因,显著性排名前3位的基因分别为Gpnmb、cd68、Caspase-1;差异共表达基因GO富集分析显示,生物过程主要涉及免疫、再生、代谢等,细胞组分包括细胞、胞外区、分子复合物等,分子功能包含受体调节活性等;KEGG通路富集分析显示,主要与CREB信号通路、神经活性配体-受体相互作用、PPAR信号通路等显著相关。RT-PCR和Western blot结果显示,与假手术组比较,模型组脊髓组织Gpnmb、cd68、Caspase-1 mRNA和蛋白表达显著升高(P<0.05);与模型组比较,中药组脊髓组织Gpnmb、cd68、Caspase-1 mRNA和蛋白表达显著降低(P<0.05)。结论益肾养髓方可能通过Gpnmb、cd68、Caspase-1等靶点治疗脊髓型颈椎病,涉及神经系统调节、免疫、代谢及反馈调节等诸多生物学过程,并通过CREB信号通路、神经活性配体-受体相互作用、PPAR信号通路等�
Objective To explore the potential mechanism of action of Yishen Yangsui Prescription in the treatment of cervical spondylotic myelopathy at the genetic and molecular levels based on RNA-Seq technology.Methods Totally 30 SD rats were randomly divided into sham-operation group,model group and TCM group,and the rat cervical spondylotic myelopathy model was established by spinal cord compression method.From one week after modeling,the TCM group was given Yishen Yangsui Prescription once a day for 8 weeks.The BBB score was used to assess the changes of motor function of the rats,the pathological changes of the spinal cord injury area were observed by transmission electron microscopy,the RNA-Seq technique was used to screen the differential genes and analyze the bioinformatics of each group,the mRNA expression level of the differential genes was detected by RT-PCR,the protein expression of differential genes was detected by Western blot.Results Compared with the sham-operation group,the BBB score in the model group was significantly lower(P<0.05).Compared with the model group,the BBB score of the TCM group significantly increased from 4 weeks after surgery(P<0.05).Transmission electron microscope observation showed that the myelin sheath of the spinal cord tissue in the model group was incomplete,broken or curled,and the rista of neurons was broken or missing,the arrangement of myelin sheath structure in the TCM group was relatively regular,and the damage to mitochondrial morphology was relatively mild.RNA-Seq technology screened 303 differentially expressed genes(153 up-regulated and 150 down-regulated)between the sham-operation group and the model group,341 differentially expressed genes(130 up-regulated and 211 down-regulated)between the model group and TCM group.The intersection of differentially expressed genes resulted in 117 co-expressed genes,and the top three genes in the significance ranking were Gpnmb,cd68 and Caspase-1,respectively.Differential co-expression gene GO enrichment analysis showed that biologic
作者
刘鹏群
银河
陈忻
张博宇
金哲峰
朱立国
LIU Pengqun;YIN He;CHEN Xin;ZHANG Boyu;JIN Zhefeng;ZHU Liguo(Beijing University of Chinese Medicine,Beijing 100029,China;Wangjing Hospital of China Academy of Chinese Medical Sciences,Beiing 100102,China;Beijing Key Laboratory of Orthopedics of Traditional Chinese Medicine,Beijing 100007,China)
出处
《中国中医药信息杂志》
CAS
CSCD
2023年第12期88-95,共8页
Chinese Journal of Information on Traditional Chinese Medicine
基金
国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-C-202003)。
关键词
益肾养髓方
脊髓型颈椎病
RNA测序技术
差异表达基因
信号通路
大鼠
Yishen Yangsui Prescription
cervical spondylotic myelopathy
RNA-Seq technique
differentially expressed genes
signaling pathway
rats
