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杨泽民,陈滢宇,袁前发,陈思羽,杨小蓉,何震宇,洪敏.脾虚证患者血清微RNA表达谱的筛选与生物信息学分析[J].Academic Journal of Second Military Medical University ,2020,41(1):99-105
脾虚证患者血清微RNA表达谱的筛选与生物信息学分析    点此下载全文 Fulltext
杨泽民1  陈滢宇2  袁前发1  陈思羽1  杨小蓉3  何震宇1  洪敏2*
1. 广东药科大学生命科学与生物制药学院生物化学与分子生物学系, 广州 510006;
2. 广东药科大学附属第一医院中医科, 广州 510080;
3. 广东药科大学附属第一医院检验科, 广州 510080
*通信作者
基金项目:国家自然科学基金青年科学基金(81102703),广东省科技计划项目(2013A032500005),广东省自然科学基金(2017A030313837),广东省中医药管理局项目(20151269),广东药学院科技处-第一医院联合自然科学培育基金项目(GYFYLH201303).
DOI:10.16781/j.0258-879x.2020.01.0099
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摘要:
      目的 筛选脾虚证患者血清miRNA表达谱进行生物信息学分析,从miRNA水平探讨脾虚证的发病机制和辨证分型规律。方法 选择高脂血症脾虚证和脾胃湿热证患者各4例,以及健康志愿者5名,提取其血清RNA,进行miRNA定量PCR芯片检测。筛选脾虚证患者血清miRNA表达谱,进行层次聚类、靶基因预测和功能注释等生物信息学分析。结果 筛选出脾虚证患者9个候选血清miRNA,其中6个表达上调,3个表达下调;候选miRNA的表达在脾虚证患者、脾胃湿热证患者及健康志愿者3组样本间存在差异。京都基因与基因组百科全书(KEGG)通路分析结果显示,6个上调miRNA调控的83个靶基因显著富集于7个通路,主要涉及细胞因子-细胞因子受体相互作用通路、病原虫感染性疾病、免疫/炎症相关的信号通路和胰腺癌等;3个下调miRNA调控的365个靶基因显著富集于5个通路,主要涉及神经营养因子信号通路、磷脂酰肌醇信号通路、RNA运输、磷酸肌醇代谢和氨基酸代谢等。结论 本研究筛选出的9个脾虚证患者候选miRNA可作为脾虚证临床辨证的潜在血清标志物,为脾虚证患者发病机制的认识和研究提供依据。
关键词:脾虚证  脾胃湿热证  高脂血症  微RNA  生物信息学
Screening and bioinformatic analysis of serum microRNA expression profiles in patients with Pi-deficiency syndrome    Fulltext
YANG Ze-min1  CHEN Ying-yu2  YUAN Qian-fa1  CHEN Si-yu1  YANG Xiao-rong3  HE Zhen-yu1  HONG Min2*
1. Department of Biochemistry and Molecular Biology, College of Life Sciences and Biopharmaceuticals, Guangdong Pharmaceutical University, Guangzhou 510006, Guangdong, China;
2. Department of Traditional Chinese Medicine, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou 510080, Guangdong, China;
3. Clinical Laboratory, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou 510080, Guangdong, China
*Corresponding author
Fund Project:Supported by National Natural Science Foundation of China for Young Scholars (81102703), Science and Technology Project of Guangdong Province (2013A032500005), Natural Science Foundation of Guangdong Province (2017A030313837), Project of Guangdong Provincial Administration of Traditional Chinese Medicine (20151269), and Natural Science Foundation for Fostering of Guangdong Pharmaceutical University (GYFYLH201303).
Abstract:
      Objective To explore the pathogenesis and syndrome differentiation of Pi-deficiency syndrome (PDS) from microRNA (miRNA) levels through screening and bioinformatic analysis of serum miRNA expression in PDS patients. Methods Four hyperlipemia patients with PDS, 4 hyperlipemia patients with Pi-Wei damp-heat syndrome (PWDS) and 5 healthy volunteers were recruited. Their serum RNA was used in miRNA quantitative PCR array experiment. Serum miRNA expression profiles in PDS patients were screened to perform bioinformatic analysis. Results Nine candidate miRNAs (6 upregulated and 3 downregulated) were screened from PDS patients. These miRNAs were able to clearly distinguish among PDS patients, PWDS patients and healthy volunteers. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed 83 target genes controlled by 6 up-regulated miRNAs were significantly enriched in 7 pathways, which were mainly involved in cytokine-cytokine receptor interaction, pathogens of infectious diseases, immune/inflammatory-related signaling pathway and pancreatic cancer; and 365 target genes controlled by 3 down-regulated miRNAs were significantly enriched in 5 pathways, which were mainly involved in signaling pathways of neurotrophin and phosphatidylinositol, RNA transport, and metabolisms of inositol phosphate and amino acid. Conclusion Our findings provide potential miRNA biomarkers for clinical syndrome differentiation of PDS patients, as well as information for understanding and studying the pathogenesis of PDS patients.
Keywords:Pi-deficiency syndrome  Pi-Wei damp-heat syndrome  hyperlipemia  microRNAs  bioinformatics
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