[1]邢 佳,曾娅莉,邓建军.肺结核患者血清外泌体中差异表达的微小RNAs及其功能分析[J].陕西医学杂志,2023,52(7):906-911，933.[doi:DOI:10.3969/j.issn.1000-7377.2023.07.030]
　XING Jia,ZENG Yali,DENG Jianjun.Differentially expressed miRNAs in serum exosomes from patients with pulmonary tuberculosis and their functional analysis[J].,2023,52(7):906-911，933.[doi:DOI:10.3969/j.issn.1000-7377.2023.07.030]
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肺结核患者血清外泌体中差异表达的微小RNAs及其功能分析
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
52
期数:
2023年7期
页码:
906-911，933
栏目:
临床检验
出版日期:
2023-07-05

文章信息/Info

Title:
Differentially expressed miRNAs in serum exosomes from patients with pulmonary tuberculosis and their functional analysis
作者:
邢 佳1; 曾娅莉1; 2; 邓建军1; 2; 3
(1.西南医科大学附属医院医学检验部,四川 泸州 646000; 2.四川绵阳四〇四医院医学检验科,四川 绵阳 621010; 3.西南科技大学,四川 绵阳 621010)
Author(s):
XING Jia; ZENG Yali; DENG Jianjun
(Department of Clinical Laboratory Science,Southwest Medical University,Luzhou 646000,China)
关键词:
微小RNAs;  肺结核;  外泌体;  信号通路;  高通量测序;  富集
Keywords:
miRNAs;  Pulmonary tuberculosis;  Exosomes;  Signaling pathway;  High-throughput sequencing;  Enrichment
分类号:
R 521
DOI:
DOI:10.3969/j.issn.1000-7377.2023.07.030
文献标志码:
A
摘要:
目的:探究肺结核患者血清外泌体中差异表达的微小RNAs(miRNAs)及其功能。方法:分子排阻法和超滤离心法提取纯化实验组及对照组血清外泌体,粒径检测、透射电镜和Western blot进行鉴定,miRNeasy Micro Kit提取外泌体总RNA; Illumina HiSeq2500高通量测序检测miRNAs表达谱; 生物信息学分析实验组和对照组差异表达的miRNAs并预测其靶基因及其靶基因潜在功能。结果:分子排阻和超滤离心获得的微囊泡直径为30～100 nm、形态完整、球形、大小较为均一,表达蛋白TSG101、HSP70、CD63、Calnexin 无表达。高通量测序分析显示,实验组显著差异表达的miRNAs共63个,其中32个上调表达,31个下调表达。GO富集分析显示差异表达的miRNAs靶基因生物学过程方面主要富集于细胞组织代谢,细胞组分方面主要富集于细胞器和细胞质,分子功能方面主要富集于催化活性、核酸结合转录因子活性。KEGG富集分析显示差异表达的miRNAs可显著富集于肾素-血管紧张素系统(RAS)、转化生长因子(TGF)、环腺苷酸单磷酸-蛋白激酶A(cGMP-PKG)等信号通路,并参与赖氨酸调节、甘油磷脂途径及类固醇激素生物合成等代谢通路和蛋白质消化吸收、神经突触囊泡循环等生物过程。结论:肺结核患者血清外泌体miRNAs表达谱与对照组比较差异具有统计学意义,差异表达的miRNAs可能通过RAS信号通路、TGF信号通路、cGMP-PKG信号通路和MAPK信号通路参与肺结核发生发展过程,在肺结核患者体内免疫调控的相关生物学功能中具有重要作用。
Abstract:
Objective:To investigate the differentially expressed microRNAs(miRNAs)in serum exosomes from patients with pulmonary tuberculosis and their functions.Methods:Serum exosomes of the experimental group and the control group were extracted and purified by molecular exclusion method and ultrafiltration centrifugation method,and identified by particle size detection,transmission electron microscopy and Western blot.Total exosomal RNA was extracted by miRNeasy Micro Kit.Illumina HiSeq2500 high-throughput sequencing was used to detect the expression profile of miRNAs.The differentially expressed miRNAs in the experimental group and the control group were analyzed by bioinformatics and their target genes and their potential functions were predicted.Results:The microvesicles obtained by molecular exclusion and ultrafiltration were 30 to 100 nm in diameter,complete in shape,spherical and uniform in size.No expression of TSG101,HSP70,CD63 and Calnexin were found.High-throughput sequencing analysis reflected that 63 miRNAs were differentially expressed in the experimental group,of which 32 were up-regulated and 31 were down-regulated.GO enrichment analysis showed that the biological processes of differentially expressed miRNAs target genes were mainly enriched in cell tissue metabolism,cell components were mainly enriched in organelles and cytoplasm,and molecular functions were enriched in dry catalytic activity and nucleic acid-binding transcription factor activity.KEGG enrichment analysis showed that differentially expressed miRNAs were significantly enriched in signaling pathways such as RAS,TGF and cGMP-PKG,and were involved in metabolic pathways such as lyoxic acid regulation,glycerophospholipid pathway and steroid hormone biosynthesis,protein digestion and absorption,and synaptic vesicle circulation.Conclusion:The expression profile of serum exosomal miRNAs in patients with pulmonary tuberculosis is significantly different from that in the control group.It is speculated that differentially expressed miRNAs may participate in the occurrence and development of pulmonary tuberculosis through RAS signaling pathway,TGF signaling pathway,CGMP-PKG signaling pathway and MAPK signaling pathway,and play an important role in the related biological functions of immune regulation in pulmonary tuberculosis patients.

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备注/Memo

备注/Memo:
基金项目:四川省科技厅重点研发项目(2019YFS0090); 四川省绵阳市卫生健康委员会科研项目(202122)
更新日期/Last Update: 2023-07-05