Table_3_MicroRNA-29c Acting on FOS Plays a Significant Role in Nonalcoholic Steatohepatitis Through the Interleukin-17 Signaling Pathway.XLSX

Nonalcoholic fatty liver disease is the most common hepatic disease in western countries and is even more ubiquitous in Asian countries. Our study determined that TH17/Treg cells were imbalanced in animal models. Based on our interest in the mechanism underlying TH17/Treg cell imbalance in nonalcoholic fatty liver mice, we conducted a joint bioinformatics analysis to further investigate this process. Common gene sequencing analysis was based on one trial from one sequencing platform, where gene expression analysis and enrichment analysis were the only analyses performed. We compared different sequencing results from different trials performed using different sequencing platforms, and we utilized the intersection of these analytical results to perform joint analysis. We used a bioinformatics analysis method to perform enrichment analysis and map interaction network analysis and predict potential microRNA sites. Animal experiments were also designed to validate the results of the data analysis based on quantitative polymerase chain reaction (qPCR) and western blotting. Our results revealed 8 coexisting differentially expressed genes (DEGs) and 7 hinge genes. The identified DEGs may influence nonalcoholic steatosis hepatitis through the interleukin-17 pathway. We found that microRNA-29c interacts with FOS and IGFBP1. Polymerase chain reaction analyses revealed both FOS and microRNA-29c expression in NASH mice, and western blot analyses indicated the same trend with regard to FOS protein levels. Based on these results, we suggest that microRNA-29c acts on FOS via the interleukin-17 signaling pathway to regulate TH17/Treg cells in NASH patients.

非酒精性脂肪性肝病（Nonalcoholic fatty liver disease）是西方国家最常见的肝脏疾病，在亚洲国家更为高发。本研究证实，动物模型中存在辅助性T细胞17（T helper 17, TH17）与调节性T细胞（regulatory T cell, Treg）失衡现象。基于我们对非酒精性脂肪性肝病小鼠模型中TH17/Treg细胞失衡机制的研究兴趣，本研究开展了联合生物信息学分析以深入探究该过程。常规基因测序分析基于单个测序平台的单次试验，仅开展了基因表达分析与富集分析。本研究对比了不同测序平台下多次试验得到的测序结果，并利用这些分析结果的交集开展联合分析。本研究采用生物信息学分析方法开展富集分析、相互作用网络绘制分析，并预测潜在的微小RNA（microRNA）结合位点。本研究还设计了动物实验，基于定量聚合酶链反应（quantitative polymerase chain reaction, qPCR）与蛋白质印迹（western blotting）技术验证数据分析结果。本研究结果显示，共筛选得到8个共存差异表达基因（differentially expressed genes, DEGs）与7个枢纽基因（hinge genes）。所鉴定出的差异表达基因可能通过白细胞介素-17（interleukin-17, IL-17）信号通路影响非酒精性脂肪性肝炎。本研究发现微小RNA-29c（microRNA-29c）可与FOS及IGFBP1相互作用。聚合酶链反应分析显示，非酒精性脂肪性肝炎小鼠模型中FOS与微小RNA-29c均存在表达；蛋白质印迹分析结果则显示，FOS蛋白水平呈现相同的变化趋势。基于上述结果，本研究提出：微小RNA-29c可通过白细胞介素-17信号通路作用于FOS，进而调控非酒精性脂肪性肝炎患者体内的TH17/Treg细胞平衡。