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目的:鉴定非酒精性脂肪性肝病(NAFLD)患者和健康对照组人血清外泌体微小RNAs(microRNAs)差异表达谱,探索miRNAs在NAFLD发病、诊断及治疗中的价值。方法:各取4例人口学特征、个人史相近的S2~3级NAFLD患者和健康对照者进行血清外泌体miRNAs高通量测序,对差异表达最显著的4条miRNAs按S1组、S2~3组、对照(Control)组每组各20例行qRT-PCR验证,并进行靶基因预测,对靶基因进行GO和KEGG富集分析。正态分布的计量资料使用n t检验或方差分析,等级资料和非正态分布资料使用秩和检验,计数资料使用Pearson n χ2检验或Fisher精确检验。n 结果:初步鉴定出差异有统计学意义(n P S1组>Control组,hsa-miR-483-3p的相对表达关系为:NAFLD组(S1或S2~3)>Control组。GO分析显示靶基因在分子功能、细胞组分、生物过程均有广泛的注释,靶基因显著富集的通路共84条,从20条与NAFLD最密切的通路中筛选出与至少10条通路都相关的靶基因共5个,即PIK3R2、AKT2、AKT3、MAPK1、NFKB1。n 结论:初步分析了NAFLD患者和健康对照组血清外泌体miRNAs差异表达谱,研究了4条miRNAs对于判断肝脂肪变性程度的价值,预测了数以千计的靶基因及其参与的复杂信号通路,为寻找无创诊断NAFLD的生物标志物和特异性治疗的新靶点提供了新的参考。“,”Objective:Differential expression of serum exosomal miRNAs were detected for NAFLD patients and healthy controls, thereby determining the role of serum exosomal miRNAs in the pathogenesis, diagnosis, and treatment of NAFLD.Methods:Four patients with S2-3 NAFLD who shared similar demographic features and personal histories, and matched healthy controls were recruited for high-throughput sequencing of serum exosomal miRNAs. Four miRNAs with the most significant differential expression were verified by qRT-PCR in three groups (S1, S2-3, and control groups) with 20 cases in each group. Target gene prediction was performed for these differentially-expressed miRNAs, along with GO and KEGG enrichment analyses for the target genes. T-test or ANOVA were used for normally distributed data. Wilcoxon rank sum test was used for ranked data and non-normally distributed data. The count data used Pearson chi-square test or Fisher\'s exact test.Results:There were 19 serum exosomal miRNAs with significantly different levels of expression (n P 2. The expression of hsa-miR-122-5p, hsa-miR-146b-5p, and hsa-miR-197-3P was highest in the S2-3 group, followed by the S1 and control groups (in order); hsa-miR-483-3p expression was higher in the NAFLD group (S1 or S2-3) than the control group. There were 84 pathways significantly enriched in target genes. From 20 pathways closely related to NAFLD, at least 5 target genes which were simultaneously correlated to all 10 pathways were screened (PIK3R2, AKT2, AKT3, MAPK1, and NFKB1).n Conclusion:Differential expression of serum exosomal miRNAs was detected in NAFLD patients and healthy controls. Four miRNAs with the greatest fold-changes were assessed to judge the severity of fatty degeneration of the liver. The research findings provide reference for non-invasive identification of new biomarkers and specific targets for NAFLD treatment.