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1.河北中医药大学 中西医结合学院,石家庄 050200
2.河北医科大学 第三医院,石家庄 050031
3.河北省离子通道功能与创新中药国际联合研究中心,石家庄 050200
Received:04 September 2025,
Revised:2025-10-31,
Accepted:24 November 2025,
Online First:27 November 2025,
Published:05 June 2026
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韩铭,张颖,孔令雅等.基于AMPK信号通路探讨血府逐瘀汤治疗代谢相关脂肪性肝病的效果及机制[J].中国实验方剂学杂志,2026,32(11):1-12.
HAN Ming,ZHANG Ying,KONG Lingya,et al.Xuefu Zhuyutang Ameliorates Metabolic-associated Fatty Liver Disease via AMPK Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(11):1-12.
韩铭,张颖,孔令雅等.基于AMPK信号通路探讨血府逐瘀汤治疗代谢相关脂肪性肝病的效果及机制[J].中国实验方剂学杂志,2026,32(11):1-12. DOI: 10.13422/j.cnki.syfjx.20251904.
HAN Ming,ZHANG Ying,KONG Lingya,et al.Xuefu Zhuyutang Ameliorates Metabolic-associated Fatty Liver Disease via AMPK Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(11):1-12. DOI: 10.13422/j.cnki.syfjx.20251904.
目的
2
采用网络药理学和动物实验相结合方法,探究血府逐瘀汤(XFZYT)治疗代谢相关脂肪性肝病(MAFLD)的作用机制。
方法
2
采用网络药理学对XFZYT关键成分、治疗MAFLD的核心靶点及信号通路进行预测;动物实验采用喂食高胆固醇饲料(HCD)4周建立MAFLD大鼠模型,给予XFZYT低剂量(2 g·kg
-1
)和XFZYT高剂量(4 g·kg
-1
)干预2周。采用生化法检测血清中天冬氨酸氨基转移酶(AST)、丙氨酸氨基转氨酶(ALT)、总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及丙二醛(MDA)、谷胱甘肽(GSH),以及肝脏中TC和TG;采用酶联免疫吸附测定法(ELISA)检测血清中白细胞介素(IL)-6、IL-1
β
、肿瘤坏死因子(TNF)-
α
水平;采用苏木素-伊红(HE)染色、油红O染色和DHE探针分别检测肝组织病理形态学、脂类沉积情况和ROS水平;蛋白免疫印迹法(Western blot)检测肝组织中腺苷酸活化蛋白激酶(AMPK)、磷酸化(p)-AMPK、核因子E
2
相关因子(Nrf2)、血红素氧合酶-1(HO-1)、核转录因子-
κ
B(NF-
κ
B)和p-NF-
κ
B的表达;采用液相色谱-串联质谱联用技术(LC-MS/MS)进行血清非靶向代谢组学分析。
结果
2
经网络药理学分析获得XFZYT治疗MAFLD的潜在靶点155个,主要核心靶点有信号转导与转录激活因子3(STAT3)、蛋白激酶B1(Akt1)、TNF和IL-6等,京都基因与基因组百科全书(KEGG)富集通路主要涉及AMPK信号通路。动物实验结果表明,与CON组比较,MOD组大鼠出现了血脂紊乱、肝功能损伤和肝内明显脂类沉积与炎症表现,血清中AST、ALT、TC、TG、LDL、MDA含量水平明显升高(
P
<
0.05),HDL、GSH含量和SOD、CAT活性明显降低(
P
<
0.05),肝脏组织中Nrf2、HO-1、p-AMPK蛋白表达水平明显降低(
P
<
0.05),p-NF-
κ
B蛋白表达水平明显升高(
P
<
0.05)。与MOD组比较,XFZYT处理组大鼠血脂紊乱、肝功能损伤明显好转,肝内脂类沉积与炎症细胞浸润明显减少,血清中AST、ALT、TC、TG、LDL、MDA水平明显降低(
P
<
0.05),HDL、GSH含量和SOD、CAT活性水平明显升高(
P
<
0.05),肝脏组织中Nrf2、HO-1、p-AMPK蛋白表达水平明显升高(
P
<
0.05),p-NF-
κ
B蛋白表达水平明显降低(
P
<
0.05)。血清代谢组学分析显示,CON组与MOD组间存在差异代谢物511种,其中231种表达上调,280种表达下调;XFZYT组与MOD组间的差异代谢物有94种,其中51种表达上调,43种表达下调;94种差异代谢产物中有11种改变最为明显,主要涉及甘油酯代谢、胆固醇代谢、胰岛素抵抗等多条与AMPK相关的通路。
结论
2
XFZYT可能通过调控AMPK信号通路对MAFLD大鼠发挥治疗作用。
Objective
2
To investigate the therapeutic mechanism of Xuefu Zhuyutang (XFZYT) for metabolic-associated fatty liver disease (MAFLD) through integrated network pharmacology and animal experiments.
Methods
2
Network pharmacology was utilized to predict the core components, key therapeutic targets, and signaling pathways of XFZYT in the treatment of MAFLD. For animal experiments, a rat model of MAFLD was established by feeding a high-cholesterol diet for 4 weeks. Intervention was then administered with low-dose (2 g·kg
-1
) and high-dose (4 g·kg
-1
) XFZYT for 2 weeks. Biochemical assays were performed to measure the serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL). In addition, the activities of superoxide dismutase (SOD) and catalase (CAT) and levels of malondialdehyde (MDA) and glutathione (GSH) in the serum were measured. The same way was adopted to measure the levels of TC and TG in the liver tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to quantify the serum levels of interleukin (IL)-6, IL-1
β
, and tumor necrosis factor-alpha (TNF-
α
). Histopathological evaluations included hematoxylin and eosin (HE) staining for liver tissue morphology, Oil Red O staining for lipid deposition, and dihydroethidium (DHE) probe staining for reactive oxygen species (ROS) levels. Western blot analysis was conducted to assess the protein levels of AMP-activated protein kinase (AMPK), phosphorylated (p)-AMPK, nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), nuclear factor-kappa B (NF-
κ
B), and p-NF-
κ
B in the liver tissue. Untargeted metabolomics analysis of the serum was performed by liquid chromatography-tandem mass spectromet
ry (LC-MS/MS).
Results
2
Network pharmacology analysis predicted 155 potential targets of XFZYT for MAFLD treatment, with core targets including signal transducer and activator of transcription 3 (STAT3), protein kinase B1 (Akt1), TNF, and IL-6. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment primarily implicated the AMPK signaling pathway. Animal experiments demonstrated that compared with the normal group, the model group exhibited dyslipidemia, hepatic function impairment, pronounced hepatic lipid deposition, and inflammatory manifestations, with elevated serum levels of AST, ALT, TC, TG, LDL, and MDA (
P
<
0.05), reduced HDL and GSH levels plus decreased SOD and CAT activities (
P
<
0.05), downregulated protein levels of Nrf2, HO-1, and p-AMPK (
P
<
0.05), and upregulated protein level of p-NF-
κ
B (
P
<
0.05) in the liver tissue. Compared with the model group, XFZYT intervention groups showed significant amelioration of dyslipidemia and hepatic function impairment, markedly reduced hepatic lipid deposition and inflammatory cell infiltration, decreased serum levels of AST, ALT, TC, TG, LDL, and MDA (
P
<
0.05), increased HDL and GSH levels plus enhanced SOD and CAT activities (
P
<
0.05), upregulated protein levels of Nrf2, HO-1, and p-AMPK (
P
<
0.05), and downregulated protein level of p-NF-
κ
B (
P
<
0.05). Serum metabolomics revealed 511 differentially expressed metabolites (231 upregulated and 280 downregulated) between normal and model groups, while XFZYT groups versus model group showed 94 differential metabolites (51 upregulated and 43 downregulated). Among them, 11 metabolites displayed the most significant alterations, with enriched pathways including glycerolipid metabolism, cholesterol metabolism, and insulin resistance, multiple of which demonstrated AMPK association.
Conclusion
2
XFZYT alleviates MAFLD by regulating the AMPK signaling pathway and associated metabolic networks.
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