目的:已知血清microRNA(miRNA)水平在非酒精性脂肪性肝病(NAFLD)中发生变化,并且可以作为有用的生物标志物。这项研究旨在全面分析所有NAFLD阶段的miRNA。
方法:我们在一个发现队列中分析了2,083个血清miRNA(183例NAFLD代表完整的NAFLD谱和10个群体对照)。通过IlluminaNextSeq测序由HTGEdgeSeq产生的miRNA文库。通过定量逆转录酶PCR,在372例其他NAFLD病例和15例人群对照中对选定的血清miRNA进行了分析。
结果:275个miRNA的水平在病例和群体对照之间存在差异。在单个NAFLD阶段中看到的差异较少,但miR-193a-5p在所有比较中均显示水平升高.相对于NAFL/非酒精性脂肪性肝炎(NASH)伴轻度纤维化(0/1期),3个miRNA(miR-193a-5p,miR-378d,和miR378d)在NASH和临床显着纤维化(2-4期)的病例中增加,7(miR193a-5p,miR-378d,miR-378e,miR-320b,miR-320c,miR-320d,和miR-320e)在NAFLD活性评分(NAS)5-8的病例中增加,而NAS较低,和3(miR-193a-5p,miR-378d,和miR-378e)增加,但1(miR-19b-3p)在脂肪变性中减少,活动,与较低的SAF活性相比,纤维化(SAF)活性评分2-4。miR-193a-5p的显著发现在患有NAFLD的额外队列中重复。对HepG2细胞的研究表明,棕榈酸处理后,miR-193a-5p表达显著下降。在病例亚组(n=80)的肝脏RNAseq数据中研究了miR-193a-5p的基因靶标;肝脏GPX8水平与血清miR-193a-5p呈正相关。
结论:血清miR-193a-5p水平与NAFLD活性分级和纤维化分期密切相关。MiR-193a-5p可能在肝脏对氧化应激的反应中起作用,并且是进行性NAFLD的潜在临床可处理的循环生物标志物。
背景:微RNA(miRNA)是可以打开或关闭基因表达的小核酸片段。这些分子可以在血液循环中检测到,在包括非酒精性脂肪性肝病(NAFLD)在内的肝病中,它们在血液中的水平可能会发生变化。看看我们是否能检测到与晚期NAFLD相关的特定miRNA,我们对183例不同严重程度的NAFLD患者和10例对照人群进行了miRNA测序.我们发现许多miRNA显示出变化,主要是增加,但1个特定的miRNAmiR-193a-5p持续增加。我们在第二组NAFLD病例中证实了这种增加。测量血液样品中的这种miRNA可能是确定患者是否患有晚期NAFLD而没有侵入性肝活检的有用方法。
OBJECTIVE: Serum microRNA (miRNA) levels are known to change in non-alcoholic fatty liver disease (NAFLD) and may serve as useful biomarkers. This study aimed to profile miRNAs comprehensively at all NAFLD stages.
METHODS: We profiled 2,083 serum miRNAs in a discovery cohort (183 cases with NAFLD representing the complete NAFLD spectrum and 10 population controls). miRNA libraries generated by HTG EdgeSeq were sequenced by Illumina NextSeq. Selected serum miRNAs were profiled in 372 additional cases with NAFLD and 15 population controls by quantitative reverse transcriptase PCR.
RESULTS: Levels of 275 miRNAs differed between cases and population controls. Fewer differences were seen within individual NAFLD stages, but miR-193a-5p consistently showed increased levels in all comparisons. Relative to NAFL/non-alcoholic steatohepatitis (NASH) with mild fibrosis (stage 0/1), 3 miRNAs (miR-193a-5p, miR-378d, and miR378d) were increased in cases with NASH and clinically significant fibrosis (stages 2-4), 7 (miR193a-5p, miR-378d, miR-378e, miR-320b, miR-320c, miR-320d, and miR-320e) increased in cases with NAFLD activity score (NAS) 5-8 compared with lower NAS, and 3 (miR-193a-5p, miR-378d, and miR-378e) increased but 1 (miR-19b-3p) decreased in steatosis, activity, and fibrosis (SAF) activity score 2-4 compared with lower SAF activity. The significant findings for miR-193a-5p were replicated in the additional cohort with NAFLD. Studies in Hep G2 cells showed that following palmitic acid treatment, miR-193a-5p expression decreased significantly. Gene targets for miR-193a-5p were investigated in liver RNAseq data for a case subgroup (n = 80); liver GPX8 levels correlated positively with serum miR-193a-5p.
CONCLUSIONS: Serum miR-193a-5p levels correlate strongly with NAFLD activity grade and fibrosis stage. MiR-193a-5p may have a role in the hepatic response to oxidative stress and is a potential clinically tractable circulating biomarker for progressive NAFLD.
BACKGROUND: MicroRNAs (miRNAs) are small pieces of nucleic acid that may turn expression of genes on or off. These molecules can be detected in the blood circulation, and their levels in blood may change in liver disease including non-alcoholic fatty liver disease (NAFLD). To see if we could detect specific miRNA associated with advanced stages of NAFLD, we carried out miRNA sequencing in a group of 183 patients with NAFLD of varying severity together with 10 population controls. We found that a number of miRNAs showed changes, mainly increases, in serum levels but that 1 particular miRNA miR-193a-5p consistently increased. We confirmed this increase in a second group of cases with NAFLD. Measuring this miRNA in a blood sample may be a useful way to determine whether a patient has advanced NAFLD without an invasive liver biopsy.