酒精及高脂饮食双重诱导下脂肪肝的转录组学变化分析

Transcriptomic analysis of fatty liver induced by alcohol and high-fat diet

目的通过酒精灌胃和高脂饮食诱导脂肪肝小鼠模型,观察小鼠肝脏表型和转录组学的改变,分析酒精、高脂饮食和联合饮食对肝脏的危害性。方法实验小鼠分为对照组(PBS)、高脂饲喂组(高脂饲料)、酒精灌胃组(40%酒精)、联合饲喂组(酒精+高脂饲料)。HE染色观察各组小鼠肝脏组织病理形态。提取肝脏RNA进行转录组测序,并针对各组转录组学的变化进行分析,观察在单一风险因素作用下和双重风险因素作用下对肝脏的影响。构建基因表达调控网络锁定网络核心基因,并对其进行验证。结果在病理层面,联合饲喂组肝脏相较于单一风险因素各组表现出更为严重的脂质累积和炎症。转录组学分析结果显示,联合饲喂组的基因表达相较于单纯的高脂饲喂组和酒精灌胃组表现出了更为明显的脂质累积通路的活跃和恶化倾向。对单风险因素和双重风险因素的差异表达基因取交集,筛选出了导致脂肪肝肝损伤的主要风险基因。功能分析结果显示高脂饮酒小鼠肝脏的基因表达变化不仅包括脂肪代谢异常,同时也和药物的不良代谢有关,最终导致肝脏炎症增加。通过网络分析可以确定Acacb,Cyp2b10和Cd36可能是高脂饮群体肝脏基因表达变化的主要靶点。结论饮酒和高脂饮食对肝脏的影响虽然都和脂肪累积引起脂肪肝相关,但并不完全一样,高脂饮食人群饮酒会对肝脏产生更严重的影响。然而关于高脂饮食和饮酒对肝脏的具体作用机制有待于进一步深入探索。

Objective To induce a mouse model of fatty liver through alcohol gavage and high-fat diet,observe the hepatic phenotype and transcriptomic changes,analyze the hepatotoxic effects of alcohol,high-fat diet,and their combinations.Methods Mice were divided into control group(PBS),high-fat diet group(high-fat feed),alcohol gavage group(40%alcohol),and combined diet group(alcohol+high-fat feed).HE staining was used to observe the pathological morphology of liver tissues in each group of mice.Liver RNA was extracted for transcriptome sequencing,and the changes in transcriptome profiles of each group were analyzed to observe the effects on the liver under the influence of a single risk factor and dual risk factors.A gene expression regulatory network was constructed to identify core genes in the network and validate them.Results At the pathological level,the combined diet group showed more severe lipid accumulation and inflammation in the liver compared to the groups with single risk factors.Transcriptome analysis results revealed that the gene expression in the combined diet group exhibited a more prominent activation of lipid accumulation pathways and a tendency towards deterioration compared to the groups with a high-fat diet alone and alcohol gavage.By intersecting the differentially expressed genes between single and double risk factor groups,key risk genes leading to liver damage in fatty liver were identified.Functional analysis results indicated that the gene expression changes in the mouse liver under high-fat alcohol conditions not only involved abnormalities in fat metabolism but also were associated with adverse drug metabolism,ultimately leading to increased liver inflammation.Network analysis identified Acacb,Cyp2b10,and Cd36 as potential key targets of gene expression changes in the liver of the high-fat diet group.Conclusion The findings of this study indicate that while both alcohol consumption and a high-fat diet contribute to fatty liver through lipid accumulation,the impact on the liver is not identical.Individuals on a high-fat diet may experience more severe liver effects when consuming alcohol.Further in-depth exploration is required to elucidate the specific mechanisms underlying the combined effects of high-fat diet and alcohol consumption on the liver.