目的 基于能量代谢与微循环探讨膈下逐瘀汤改善代谢相关脂肪性肝炎（MASH）的作用机制。方法 选取8 dpf斑马鱼，随机分为对照组、模型组、洛伐他汀（阳性药，4 μg·mL^-1）组和膈下逐瘀汤低、中、高质量浓度（20、50、80 μg·mL^-1）组，除对照组外，使用4%浓度的果糖溶液建立MASH模型，每6小时更换药液，连续给药72 h。观察水质中残留的饲料以判断各组斑马鱼进食情况，观察斑马鱼幼鱼存活率、体长；苏木精-伊红（HE）染色观察肝脏组织形态；油红O染色观察肝脏面积及脂质沉积情况；制备斑马鱼匀浆上清液，ELISA试剂盒检测总胆固醇（TC）、三酰甘油（TG）、丙酮酸、葡萄糖、腺嘌呤核苷三磷酸（ATP）酶、血管内皮生长因子（VEGF）、血管生成素1（Ang-1）、Ang-2；微循环仪下观察并记录尾部微血管的血流速度变化；Western blotting检测沉寂信息调节因子（SIRT1）、AMP依赖的蛋白激酶（AMPK）、p-AMPK蛋白的表达水平。结果 与模型组相比，膈下逐瘀汤组斑马鱼存活率呈上升趋势但无明显差异，体长和体质量明显下降（P＜0.05、0.01）；斑马鱼肝脏面积显著减小（P＜0.05、0.01），脂滴明显减少，染色明显减轻，肝内脂质沉积减少；肝细胞脂肪变性和气球样变性得到明显改善；TC、TG水平显著下降（P＜0.05、0.01）；丙酮酸含量明显增加（P＜0.05、0.01），葡萄糖含量显著减少（P＜0.05、0.01），ATP酶活力水平明显增加（P＜0.05、0.01），VEGF、Ang-1水平明显降低（P＜0.05、0.01），Ang-2水平明显升高（P＜0.05、0.01），斑马鱼各部位血流速度显著增加（P＜0.05、0.01），斑马鱼SIRT1、AMPK、p-AMPK蛋白的表达水平明显上升（P＜0.05、0.01）。结论 膈下逐瘀汤可能通过改善微循环和激活AMPK/SIRT1信号通路来改善MASH斑马鱼的能量代谢，进而减轻MASH引起的肝损伤。

Objective To investigate the mechanism of Gexia Zhuyu Decoction in ameliorating metabolic-associated steatohepatitis (MASH) through modulating energy metabolism and microcirculation.Methods Zebrafish at 8 days post-fertilization (dpf) were randomly divided into a control group, a model group, a lovastatin (positive drug, 4 μg·mL^-1) group, and low-, medium-, and highconcentration groups of Gexia Zhuyu Decoction (20, 50, and 80 μg·mL^-1). Except for the control group, a 4% fructose solution was used to establish a MASH model. The drug solution was changed every 6 h and administered continuously for 72 h. The remaining feed in the water was observed to determine the feeding status of zebrafish in each group. The survival rate and body length of zebrafish larvae were observed. HE staining was used to observe the morphology of liver tissue. Oil Red O staining was used to observe the liver area and lipid deposition. Zebrafish homogenate supernatant was prepared, and ELISA kits were used to detect total cholesterol (TC), triglycerides (TG), pyruvate, glucose, adenine nucleoside triphosphate (ATP) enzyme, vascular endothelial growth factor (VEGF), angiopoietin 1 (Ang-1), and Ang-2. The blood flow velocity changes in the tail microvessels were observed and recorded under a microcirculation instrument. Western blotting was used to detect the expression levels of silent information regulator 1 (SIRT1), AMPactivated protein kinase (AMPK), and p-AMPK proteins.Results Compared with the model group, the survival rate of zebrafish in the Gexia Zhuyu Decoction group showed an upward trend but no significant difference. The body length and body weight of zebrafish decreased significantly (P<0.05, 0.01). The liver area of zebrafish decreased significantly (P<0.05, 0.01), the lipid droplets decreased significantly, the staining was significantly reduced, and the lipid deposition in the liver was reduced. The fatty degeneration and ballooning degeneration of liver cells were significantly improved. The levels of TC and TG decreased significantly (P<0.05, 0.01). The content of pyruvate increased significantly (P<0.05, 0.01), the content of glucose decreased significantly (P<0.05, 0.01), the activity level of ATPase increased significantly (P<0.05, 0.01), the levels of VEGF and Ang-1 decreased significantly (P<0.05, 0.01), the level of Ang-2 increased significantly (P<0.05, 0.01), and the blood flow velocity in various parts of zebrafish increased significantly (P<0.05, 0.01). The expression levels of SIRT1, AMPK, and p-AMPK proteins in zebrafish increased significantly (P<0.05, 0.01).Conclusion Gexia Zhuyu Decoction may improve the energy metabolism of MASH zebrafish by improving microcirculation and activating the AMPK/SIRT1 signaling pathway, thereby alleviating liver damage caused by MASH.

R285.5

国家自然科学基金项目（81603418）；黑龙江优秀青年人才项目（2020YQ05）；黑龙江中医药大学研究生创新科研项目（2024yjscx124）