Background

Nonalcoholic fatty liver disease and its advanced stage, nonalcoholic steatohepatitis (NASH), are the major cause of hepatocellular carcinoma (HCC) and other end-stage liver disease. However, the potential mechanism and therapeutic strategies have not been clarified. This study aimed to identify potential roles of miRNA/mRNA axis in the pathogenesis and drug combinations in the treatment of NASH.

Methods

Microarray GSE59045 and GSE48452 were downloaded from the Gene Expression Omnibus and analyzed using R. Then we obtained differentially expressed genes (DE-genes). DAVID database was used for Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathway analysis. Protein-protein interaction (PPI) networks were used for the identification of hub genes. We found upstream regulators of hub genes using miRTarBase. The expression and correlation of key miRNA and its targets were detected by qPCR. Drug Pair Seeker was employed to predict drug combinations against NASH. The expression of miRNA and hub genes in HCC was identified in the Cancer Genome Atlas database and Human Protein Atlas database.

Results

Ninety-four DE-genes were accessed. GO and KEGG analysis showed that these predicted genes were linked to lipid metabolism. Eleven genes were identified as hub genes in PPI networks, and they were highly expressed in cells with vigorous lipid metabolism. hsa-miR-335-5p was the upstream regulator of 9 genes in the 11 hub genes, and it was identified as a key miRNA. The hub genes were highly expressed in NASH models, while hsa-miR-335-5p was lowly expressed. The correlation of miRNA-mRNA was established by qPCR. Functional verification indicated that hsa-miR-335-5p had inhibitory effect on the development of NASH. Finally, drug combinations were predicted and the expression of miRNA and hub genes in HCC was identified.

Conclusions

In the study, potential miRNA-mRNA pathways related to NASH were identified. Targeting these pathways may be novel strategies against NASH.

中文翻译：

---

非酒精性脂肪性肝炎的关键因素和潜在药物组合：生物信息学分析和基于实验验证的研究

背景

非酒精性脂肪性肝病及其晚期非酒精性脂肪性肝炎 (NASH) 是肝细胞癌 (HCC) 和其他终末期肝病的主要原因。然而，潜在的机制和治疗策略尚未阐明。本研究旨在确定 miRNA/mRNA 轴在 NASH 的发病机制和药物组合治疗中的潜在作用。

方法

从 Gene Expression Omnibus 下载微阵列 GSE59045 和 GSE48452 并使用 R 进行分析。然后我们获得了差异表达基因 (DE-genes)。DAVID 数据库用于基因本体论 (GO) 分析和京都基因和基因组百科全书 (KEGG) 富集途径分析。蛋白质-蛋白质相互作用（PPI）网络用于鉴定中心基因。我们使用 miRTarBase 发现了中枢基因的上游调节因子。qPCR检测关键miRNA及其靶标的表达和相关性。Drug Pair Seeker 用于预测针对 NASH 的药物组合。在癌症基因组图谱数据库和人类蛋白质图谱数据库中鉴定了 HCC 中 miRNA 和中枢基因的表达。

结果

访问了 94 个 DE 基因。GO和KEGG分析表明，这些预测基因与脂质代谢有关。11个基因被鉴定为PPI网络中的枢纽基因，它们在脂质代谢旺盛的细胞中高度表达。hsa-miR-335-5p是11个hub基因中9个基因的上游调控因子，被鉴定为关键miRNA。枢纽基因在 NASH 模型中高表达，而 hsa-miR-335-5p 低表达。通过qPCR建立miRNA-mRNA的相关性。功能验证表明hsa-miR-335-5p对NASH的发展具有抑制作用。最后，预测了药物组合，并确定了 HCC 中 miRNA 和中枢基因的表达。

结论

在这项研究中，确定了与 NASH 相关的潜在 miRNA-mRNA 通路。针对这些途径可能是针对 NASH 的新策略。