基于斑马鱼模型和网络药理学研究小柴胡汤治疗非酒精性脂肪肝病的作用及潜在机制

The Effect and Potential Mechanism of Xiaochaihu Decoction on Non-alcoholic Fatty Liver Disesease Based on Zebrafish Model and Network Pharmacology

关键词:  非酒精性脂肪肝  小柴胡汤  斑马鱼  网络药理学

Key words:  nonalcoholic fatty liver disease  Xiaochaihu Decoction  zebrafish  network pharmacology

摘要:

目的 基于斑马鱼模型及网络药理学技术探究小柴胡汤治疗非酒精性脂肪肝病(NAFLD)的活性成分、作用靶点及潜在机制。方法 采用高脂高胆固醇饲料喂养诱导建立斑马鱼幼鱼非酒精性脂肪肝病模型,以斑马鱼肝脏表型、H E病理切片和炎症因子表达水平等作为评价指标,综合评价小柴胡汤对非酒精性脂肪肝的作用,结合网络药理学分析其可能的作用机制。结果 与模型组相比,小柴胡汤可降低斑马鱼幼鱼的体质量与BMI指数增加,缓解肝脏脂肪变性、脂质沉积和结构损伤,剂量依赖性地降低斑马鱼体内的TG、TC、ALT、AST含量,降低ROS水平并提高SOD酶活力,同时降低斑马鱼体内的炎症因子TNF-α、IL-6、IL-1β的mRNA表达水平。网络药理学得到小柴胡汤药效活性成分198个与作用靶点108个,主要富集于PI3K-Akt信号通路及细胞凋亡相关信号通路, PPI网络分析得到汉黄芩素、6-姜辣素等关键活性成分和PTGS2等核心靶点,高剂量小柴胡汤可以显著调节NAFLD斑马鱼体内上述核心靶点基因的mRNA表达水平。结论 小柴胡汤可通过多成分、多途径、多靶点干预NAFLD早期的NAFL-NASH进展,其主要活性成分汉黄芩素等可通过作用于PTGS2、AKT1等靶点,参与调控PI3K-Akt信号通路及细胞凋亡相关信号通路等来发挥降脂保肝抗氧化抗炎的作用。

Abstract:

ABSTRACT: OBJECTIVE To explore the active ingredients, targets, and potential mechanisms of Xiaochaihu Decoction (XCHD) in the treatment of non-alcoholic fatty liver disease (NAFLD) based on the zebrafish model and network pharmacology technology. METHORDS The zebrafish NAFLD model was established by feeding zebrafish larvae with high-fat and high-cholesterol diet. The evaluation indicators, including zebrafish liver phenotype, H&E pathological slices, and expression levels of inflammatory factors, were used to comprehensively evaluate the effect of XCHD on NAFLD. Network pharmacology analysis was used to explore its potential mechanisms of action. RESULTS Compared with the model group, XCHD could reduce the body weight and BMI index of zebrafish larvae, alleviate liver steatosis, lipid deposition, and structural damage, dose-dependently reduce the levels of TG, TC, ALT, AST in zebrafish, decrease ROS levels, and increase SOD enzyme activity. It also reduced the mRNA expression levels of inflammatory factors TNF-α, IL-6, IL-1β in zebrafish. 198 active ingredients and 108 targets of XCHD was identified by Network pharmacology, mainly enriched in the PI3K-Akt signaling pathway and apoptosis-related signaling pathways. PPI network analysis revealed key active ingredients such as baicalin, 6-gingerol, and core targets such as PTGS2. High-dose XCHD significantly regulated the mRNA expression levels of these core targets in NAFLD zebrafish. CONCLUSION XCHD can intervene in the early progression of NAFL to NASH through multiple components, pathways, and targets. Its main active ingredient, baicalin, can exert lipid-lowering, hepatoprotective, antioxidant, and anti-inflammatory effects by acting on targets such as PTGS2 and AKT1, and participating in the regulation of the PI3K-Akt signaling pathway and apoptosis-related signaling pathways.