目的:小儿非酒精性脂肪性肝病(NAFLD)是一个日益严重的问题,但它的潜在机制知之甚少。我们使用转录组报告细胞测定法来研究患有和不患有NAFLD的儿童的血清在肝细胞报告细胞中诱导的转录特征的差异。
方法:我们研究了45名NAFLD儿童和28名非NAFLD儿童的血清样本。血清用于诱导培养的HepaRG细胞中的基因表达,RNA测序用于确定基因表达。使用计算技术来比较基因表达模式。
结果:来自NAFLD儿童的血清诱导了195个基因的表达,这些基因在肝细胞中的表达与肥胖对照组相比有显著差异。NAFLD与促进炎症的基因表达增加有关,胶原蛋白合成,和细胞外基质重塑。此外,参与内源性和外源性生物代谢的基因表达较低,和过氧化物酶体功能的下调,氧化磷酸化,和异种生物,胆汁酸,和脂肪酸代谢。一个13个基因的签名,包括TREM1和MMP1的上调和CYP2C9的下调,均与小儿NAFLD的所有诊断类别相关.
结论:NAFLD患儿血清的细胞外环境可诱导肝细胞报告系统区分的特定基因谱。循环因素可能有助于小儿NAFLD中的炎症和细胞外基质重塑,并损害异种生物和内源性代谢。
OBJECTIVE: Pediatric nonalcoholic fatty liver disease (NAFLD) is a growing problem, but its underlying mechanisms are poorly understood. We used transcriptomic reporter cell assays to investigate differences in transcriptional signatures induced in hepatocyte reporter cells by the sera of children with and without NAFLD.
METHODS: We studied serum samples from 45 children with NAFLD and 28 children without NAFLD. The sera were used to induce gene expression in cultured HepaRG cells and RNA-sequencing was used to determine gene expression. Computational techniques were used to compare gene expression patterns.
RESULTS: Sera from children with NAFLD induced the expression of 195 genes that were significantly differentially expressed in hepatocytes compared to controls with obesity. NAFLD was associated with increased expression of genes promoting inflammation, collagen synthesis, and extracellular matrix remodeling. Additionally, there was lower expression of genes involved in endobiotic and xenobiotic metabolism, and downregulation of peroxisome function, oxidative phosphorylation, and xenobiotic, bile acid, and fatty acid metabolism. A 13-gene signature, including upregulation of TREM1 and MMP1 and downregulation of CYP2C9, was consistently associated with all diagnostic categories of pediatric NAFLD.
CONCLUSIONS: The extracellular milieu of sera from children with NAFLD induced specific gene profiles distinguishable by a hepatocyte reporter system. Circulating factors may contribute to inflammation and extracellular matrix remodeling and impair xenobiotic and endobiotic metabolism in pediatric NAFLD.