目的 研究灵芝与葛根合用对慢性氧化应激诱导高血压的协同降压效果及其作用机制。方法 运用网络药理学获取灵芝、葛根有效成分通过调控氧化应激抗高血压的靶点，通过蛋白质-蛋白质相互作用（PPI）网络分析核心靶点；对核心靶点进行京都基因与基因组百科全书（KEGG）通路和基因本体（GO）富集分析。构建L-NG-硝基精氨酸甲酯（L-NAME）诱导的高血压大鼠模型，将56只高血压模型大鼠随机分为模型组，灵芝低、中、高剂量(50、100、300 mg·kg^-1)组，葛根低、中、高剂量(50、100、300 mg·kg^-1)组，并随机选取8只对照大鼠作对照组，各给药组每天ig给予药材研磨成的超细粉末1次，连续给药3周，检测给药3周后各组收缩压（SBP），筛选后续合用给药剂量。将48只高血压模型大鼠随机分为灵芝(100 mg·kg^-1)组、葛根(50 mg·kg^-1)组、灵芝(100 mg·kg^-1)-葛根(50 mg·kg^-1)组、卡托普利(Capt,10 mg·kg^-1)组、模型组，并随机选取8只对照大鼠作对照组，连续给药3周，每周测量1次体质量及SBP；采用ELISA试剂盒检测血清血管紧张素Ⅱ（AngⅡ）、超氧化物歧化酶（SOD）、丙二醛（MDA）水平及胸主动脉组织血管细胞黏附分子-1（VCAM-1）、总抗氧化能力（T-AOC）、前列环素2（PGI2）浓度；称量心脏质量，计算心脏指数；苏木素-伊红（HE）染色检测胸主动脉组织形态变化；Western blotting法检测胸主动脉组织核因子-相关因子2（Nrf2）、上游调控蛋白Kelch样ECH关联蛋白1（Keap1）及下游靶蛋白血红素加氧酶1（HO-1）、单核苷酸氧化酶1（NQO1）的表达水平，以反映氧化应激通路变化。结果 网络药理学分析结果显示，灵芝和葛根鉴定30个与高血压和氧化应激相关的共同靶点，KEGG通路分析对其显著富集“流体剪切应力和动脉粥样硬化”和“PI3K-Akt”信号通路，GO富集分析对其显著富集“对氧化应激的反应”等生物学功能。体内实验证实，与模型组相比，灵芝和葛根可协同增效显著增加模型大鼠体质量、降低SBP，降低血清AngⅡ、血清MDA含量，心脏指数及胸主动脉VCAM-1浓度，并提高血清SOD及胸主动脉NO、T-AOC、PGI2水平（P<0.05、0.01、0.001）；明显改善胸主动脉损伤；显著下调Keap1蛋白表达，上调Nrf2、HO-1及NQO1蛋白表达水平（P<0.05、0.01、0.001）。结论 灵芝与葛根通过“流体剪切应力和动脉粥样硬化”和PI3K-Akt信号通路，共同调控Keap1-Nrf2-NQO1/HO-1通路增强抗氧化应激能力，对慢性氧化应激诱导的高血压模型起到协同增效的作用。

Objective To investigate the synergistic antihypertensive effects and underlying mechanisms of Ganoderma lucidum combined with Pueraria lobata in treating chronic oxidative stress-induced hypertension, providing a reference for developing foodmedicine dual-use Chinese herbal medicines for hypertension prevention and treatment. Methods Network pharmacology was used to obtain the intersection of the effective components of G. lucidum and P. lobata with the targets of hypertension and oxidative stress diseases, and the core targets were analyzed through protein network interaction. The core targets were subjected to Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway and Gene Ontology（GO） enrichment analysis. A rat model of hypertension induced by LNG-nitroarginine methyl ester（L-NAME） was established. Fifty-six hypertensive model rats were randomly and evenly divided into the model group, low-, medium-, and high-dose G. lucidum groups(50, 100, and 300 mg·kg^-1), and low-, medium-, and high-dose P. lobata groups(50, 100, and 300 mg·kg^-1), and eight control rats were randomly selected as the control group. Each drug group was given a single ig administration of the ultrafine powder of the ground herbs once a day for three consecutive weeks. The systolic blood pressure（SBP） of each group was measured after three weeks of administration to screen the subsequent combined administration doses. Forty-eight hypertensive model rats were randomly and evenly divided into the G. lucidum(100 mg·kg^-1) group, the P. lobata(50 mg·kg^-1) group, the G. lucidum(100 mg·kg^-1)-P. lobata(50 mg·kg^-1) group, the captopril(Capt, 10 mg·kg^-1) group, and the model group, and eight control rats were randomly selected as the control group. The rats were given drugs for three consecutive weeks, and the body weight and SBP were measured once a week. The levels of serum angiotensin II（Ang II）, superoxide dismutase（SOD）, and malondialdehyde（MDA） and the concentrations of vascular cell adhesion molecule-1（VCAM-1）, total antioxidant capacity（T-AOC）, and prostacyclin 2（PGI2） in the thoracic aorta tissue were detected using ELISA kits. The heart weight was measured, and the heart index was calculated. The morphological changes of the thoracic aorta tissue were detected by hematoxylin-eosin（HE） staining. The expression levels of nuclear factor erythroid 2-related factor 2（Nrf2）, upstream regulatory protein Keap1, and downstream target proteins heme oxygenase 1（HO-1） and quinone oxidoreductase 1（NQO1） in the thoracic aorta tissue were detected by Western blotting to reflect the changes in the oxidative stress pathway. Results The results of network pharmacology analysis showed that G. lucidum and P. lobata identified 30 common targets related to hypertension and oxidative stress. KEGG pathway analysis significantly enriched "Fluid shear stress and atherosclerosis" and "PI3K-Akt" signaling pathways for them, and GO enrichment analysis significantly enriched "Response to oxidative stress" and other biological functions for them. In vivo experiments confirmed that compared with the model group, G. lucidum and P. lobata could synergistically enhance significantly body weight, reduce SBP, reduce serum AngⅡ, serum MDA content, cardiac index and thoracic aortic VCAM-1 concentration, and increase serum SOD, thoracic aortic NO, T-AOC and PGI2 levels（P < 0.05, 0.01, 0.001）; Significantly improve thoracic aortic injury; significantly down-regulate Keap1 protein expression, and up-regulate Nrf2, HO-1 and NQO1 protein expression levels（P < 0.05, 0.01, 0.001）. Conclusion G. lucidum and P. lobata synergistically enhance antioxidant stress capacity by jointly regulating the Keap1-Nrf2-NQO1/HO-1 pathway through the “fluid shear stress and atherosclerosis” and PI3K-AKT signaling pathways, thereby exerting a synergistic effect on the chronic oxidative stress-induced hypertension model.

R285

中央支持地方高校改发展资金人才培养项目; 2024年种业创新发展资金项目; 黑龙江省高校2024年基本科研业务费专项资金(2024-KYYWF-1021); 哈尔滨商业大学2022年博士科研支撑计划(22BQ59)