心力衰竭患者血浆外泌体microRNA检测及其功能分析
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摘要
目的明确心力衰竭(心衰)患者血浆外泌体microRNA(miRNA)表达谱的差别,寻找其在心衰早期诊断和预后评估中的价值。方法应用外泌体提取试剂盒抽提10例心衰患者及10例非心衰对照者外周血浆外泌体,对血浆外泌体miRNA进行高通量测序,初步筛选差异表达miRNA,并通过实时荧光定量PCR方法验证,分析差异表达miRNA的功能及其与NT-proBNP的相关性。结果 miRNA高通量测序共筛选出24条显著差异表达的血浆外泌体miRNA。其中14条miRNA表达显著上调,10条miRNA表达明显下调。定量PCR验证和高通量测序筛选结果一致。差异表达miRNA的功能涉及炎性反应、细胞凋亡、增值、自噬等。相关性分析提示miR-122-5p与NT-proBNP具有相关性(r=0.537,P=0.015)。结论血浆外泌体来源miRNA可能成为潜在的辅助诊断心力衰竭和评估预后的一类新型生物标志物。
Objective To determine the difference in expression profiles of plasma exosomal microRNA(miRNA) in patients with heart failure(HF), and find out the value of exosomal miRNA in reviewing on early diagnosis and prognosis of HF. Methods The samples of peripheral plasma exosome were extracted from HF patients(n=10) and non-HF patients(n=10) by using exosome extract kit, and then high-throughput sequencing technique was applied for plasma exosomal miRNA. The miRNA with different expressions were initially screened and identified by using RT-qPCR. The correlation between function of miRNA with different expressions and N-terminal pro-brain natriuretic peptide(NT-proBNP) was analyzed. Results There were totally 24 plasma exosomal miRNA screened by using high-throughput sequencing technique. Among them 14 plasma exosomal miRNA had up-regulated expressions and 10 had down-regulated expressions. The results of RT-qPCR were consistent with those of high-throughput sequencing technique. The functions of plasma exosomal miRNA with different expressions involved inflammatory reactions, apoptosis, proliferation and autophagy. The correlation analysis showed that miR-122-5 p was correlated to NT-proBNP(r=0.537, P=0.015). Conclusion The plasma exosomal miRNA may be a new type of biomarker for auxiliary diagnosis and prognosis review of HF.
Objective To determine the difference in expression profiles of plasma exosomal microRNA(miRNA) in patients with heart failure(HF), and find out the value of exosomal miRNA in reviewing on early diagnosis and prognosis of HF. Methods The samples of peripheral plasma exosome were extracted from HF patients(n=10) and non-HF patients(n=10) by using exosome extract kit, and then high-throughput sequencing technique was applied for plasma exosomal miRNA. The miRNA with different expressions were initially screened and identified by using RT-qPCR. The correlation between function of miRNA with different expressions and N-terminal pro-brain natriuretic peptide(NT-proBNP) was analyzed. Results There were totally 24 plasma exosomal miRNA screened by using high-throughput sequencing technique. Among them 14 plasma exosomal miRNA had up-regulated expressions and 10 had down-regulated expressions. The results of RT-qPCR were consistent with those of high-throughput sequencing technique. The functions of plasma exosomal miRNA with different expressions involved inflammatory reactions, apoptosis, proliferation and autophagy. The correlation analysis showed that miR-122-5 p was correlated to NT-proBNP(r=0.537, P=0.015). Conclusion The plasma exosomal miRNA may be a new type of biomarker for auxiliary diagnosis and prognosis review of HF.
引文
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[2]黄曦乐,李展业,郭嘉琪.人参中药制剂治疗冠心病心绞痛的Meta分析[J].实用医学杂志,2015,31(3):454-7.
[3]毕颖斐,毛静远,王贤良,等.中医药防治冠心病临床优势及有关疗效评价的思考[J].中医杂志,2015,56(5):437-40.
[4]唐梅森,黄政德,向忠军,等.加味丹参饮联合西药治疗冠心病心绞痛心血瘀阻证68例疗效观察[J].中医杂志,2015,56(5):395-7.
[5]Singer RE,Moss K,Kim SJ,et al. Oxidative Stress and IgG Antibody Modify Periodontitis-CRP Association[J]. Journal of Dental Research, 2015,94(12):1698.
[6]Tachezy M,Anusic I,Rothenh fer S,et al.[Erratum to:Perioperative CRP quantification for appendectomy:Clinically useful or a waste of money?][J]. Chirurg,2015,86(9):1-8.
[7]薛静.血必净注射液对伴心肌损害重症肺炎患者sICAM-1、hs-CRP、IL-17及心肌损伤指标的影响[J].现代中西医结合杂志,2016,25(35):3909-12.
[8]许震,孙生安,刘帅峰,等.左氧氟沙星不同用药方法对急性化脓性阑尾炎术后切口感染及血清sICAM-1、IL-8、TNF-a的影响研究[J].中华医院感染学杂志,2016, 26(22):5169-71.
[9]Cai A,Li G,Chen J,et al. Increased serum level of Lp-PLA2 is independently associated with the severity of coronary artery diseases:a cross-sectional study of Chinese population[J]. BMC Cardiovascular Disorders,2015,15(1):14.
[10]路亚枫,吕树铮,陈韵岱,等.冠心病临界病变患者血管因子与冠脉斑块形态学特征的相关性研究[J].中华急诊医学杂志,2015,24(3):267-72.