目的 基于能量代謝與微循環(huán)探討膈下逐瘀湯改善代謝相關(guān)脂肪性肝炎（MASH）的作用機(jī)制。方法 選取8 dpf斑馬魚，隨機(jī)分為對照組、模型組、洛伐他汀（陽性藥，4 μg·mL^-1）組和膈下逐瘀湯低、中、高質(zhì)量濃度（20、50、80 μg·mL^-1）組，除對照組外，使用4%濃度的果糖溶液建立MASH模型，每6小時更換藥液，連續(xù)給藥72 h。觀察水質(zhì)中殘留的飼料以判斷各組斑馬魚進(jìn)食情況，觀察斑馬魚幼魚存活率、體長；蘇木精-伊紅（HE）染色觀察肝臟組織形態(tài)；油紅O染色觀察肝臟面積及脂質(zhì)沉積情況；制備斑馬魚勻漿上清液，ELISA試劑盒檢測總膽固醇（TC）、三酰甘油（TG）、丙酮酸、葡萄糖、腺嘌呤核苷三磷酸（ATP）酶、血管內(nèi)皮生長因子（VEGF）、血管生成素1（Ang-1）、Ang-2；微循環(huán)儀下觀察并記錄尾部微血管的血流速度變化；Western blotting檢測沉寂信息調(diào)節(jié)因子（SIRT1）、AMP依賴的蛋白激酶（AMPK）、p-AMPK蛋白的表達(dá)水平。結(jié)果 與模型組相比，膈下逐瘀湯組斑馬魚存活率呈上升趨勢但無明顯差異，體長和體質(zhì)量明顯下降（P＜0.05、0.01）；斑馬魚肝臟面積顯著減?。?i>P＜0.05、0.01），脂滴明顯減少，染色明顯減輕，肝內(nèi)脂質(zhì)沉積減少；肝細(xì)胞脂肪變性和氣球樣變性得到明顯改善；TC、TG水平顯著下降（P＜0.05、0.01）；丙酮酸含量明顯增加（P＜0.05、0.01），葡萄糖含量顯著減少（P＜0.05、0.01），ATP酶活力水平明顯增加（P＜0.05、0.01），VEGF、Ang-1水平明顯降低（P＜0.05、0.01），Ang-2水平明顯升高（P＜0.05、0.01），斑馬魚各部位血流速度顯著增加（P＜0.05、0.01），斑馬魚SIRT1、AMPK、p-AMPK蛋白的表達(dá)水平明顯上升（P＜0.05、0.01）。結(jié)論 膈下逐瘀湯可能通過改善微循環(huán)和激活A(yù)MPK/SIRT1信號通路來改善MASH斑馬魚的能量代謝，進(jìn)而減輕MASH引起的肝損傷。

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

國家自然科學(xué)基金項目（81603418）；黑龍江優(yōu)秀青年人才項目（2020YQ05）；黑龍江中醫(yī)藥大學(xué)研究生創(chuàng)新科研項目（2024yjscx124）