Chen Bo, Zheng Yang-Min, Zhang Miao-Qing, Han Ying, Zhang Jing-Pu, Hu Chang-Qin
Key Laboratory of Biotechnology of Antibiotics, The National Health Commission (NHC), Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Beijing Key Laboratory of Antimicrobial Agents, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Front Pharmacol. 2020 Jan 8;10:1504. doi: 10.3389/fphar.2019.01504. eCollection 2019.
The prevalence of non-alcohol fatty liver disease (NAFLD) is increasing in children and adolescents who are mostly resulted from overfeeding. Previous studies demonstrate that berberine (BBR), a compound derived from plant, has beneficial effects on NAFLD in adults but poorly understood in the pediatric population. This study employed a larval zebrafish model to mimic the therapeutic effects of BBR in the pediatric population and the mechanisms underlying its hepatoprotection. High-cholesterol diet (HCD)-fed zebrafish exposed to BBR at doses of 0, 1, 5, and 25 μM. After the larvae were treated with BBR for 10 days, its effect on hepatic steatosis was evaluated. We introduced Raman imaging and three-dimensional (3D) molecular imaging to detect changes in the biochemical composition and reactive oxygen species (ROS) levels of zebrafish liver. Gene expression microarray was performed to identify differentially expressed genes (DEGs) followed by gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and functional category analysis. BBR (5 and 25 μM) administration prevented HCD-induced liver lipid accumulation in larval zebrafish. The result was further confirmed by the pathological observation. Raman mapping indicated that the biochemical composition in the liver of BBR-treated group shifted to the control. The quantitative analysis of 3D imaging showed that the ROS level was significantly decreased in the liver of BBR-treated larvae. In the livers of the BBR group, we found 468 DEGs, including 172 genes with upregulated expression and 296 genes with downregulated expression. Besides, GO enrichment, KEGG pathway, and functional category analysis showed that various processes related to glucolipid metabolism, immune response, DNA damage and repair, and iron were significantly enriched with DEGs. The expression levels of the crucial genes from the functional analysis were also confirmed by quantitative PCR (qPCR). BBR can significantly improve hepatic steatosis in HCD-fed zebrafish larvae. Its mechanisms might be associated with the regulation of lipid metabolism, oxidative stress, and iron homeostasis. Raman imaging in larval zebrafish might become a useful tool for drug evaluation. Mainly, the gene expression profiles provide molecular information for BBR on the prevention and treatment of pediatric NAFLD.
非酒精性脂肪性肝病（NAFLD）在儿童和青少年中的患病率正在上升，这主要是由过度喂养导致的。先前的研究表明，黄连素（BBR）是一种从植物中提取的化合物，对成人的NAFLD有有益作用，但在儿科人群中的了解较少。本研究采用幼体斑马鱼模型来模拟BBR在儿科人群中的治疗效果及其肝脏保护的潜在机制。将高胆固醇饮食（HCD）喂养的斑马鱼暴露于0、1、5和25 μM剂量的BBR中。在用BBR处理幼虫10天后，评估其对肝脂肪变性的影响。我们引入拉曼成像和三维（3D）分子成像来检测斑马鱼肝脏生化组成和活性氧（ROS）水平的变化。进行基因表达微阵列以鉴定差异表达基因（DEG），随后进行基因本体（GO）、京都基因与基因组百科全书（KEGG）通路和功能类别分析。给予BBR（5和25 μM）可预防HCD诱导的幼体斑马鱼肝脂质积累。病理观察进一步证实了这一结果。拉曼映射表明，BBR处理组肝脏中的生化组成向对照组转变。3D成像的定量分析表明，BBR处理的幼虫肝脏中的ROS水平显著降低。在BBR组的肝脏中，我们发现了468个DEG，包括172个表达上调的基因和296个表达下调的基因。此外，GO富集、KEGG通路和功能类别分析表明，与糖脂代谢、免疫反应、DNA损伤和修复以及铁相关的各种过程在DEG中显著富集。功能分析中关键基因的表达水平也通过定量PCR（qPCR）得到了证实。BBR可以显著改善HCD喂养的斑马鱼幼虫的肝脂肪变性。其机制可能与脂质代谢、氧化应激和铁稳态的调节有关。幼体斑马鱼中的拉曼成像可能成为药物评估的有用工具。主要是，基因表达谱为BBR预防和治疗儿科NAFLD提供了分子信息。
