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| 丹知青娥方通过PCSK9/LDLR通路降脂作用机制研究 |
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宋俊丽1, 师辉辉1, 陈广侠2, 黄宇星3, 毛浩萍1, 马尚伟3
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1.天津中医药大学, 天津 301617;2.天津中医药大学第一附属医院, 天津 300381;3.天津中医药大学第二附属医院, 天津 300250
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| 摘要: |
| [目的] 基于网络药理学、分子对接和实验探究丹知青娥方（DZQEF）降脂的作用机制。[方法] 检测丹知青娥方对HepG2细胞红色荧光标记低密度脂蛋白（Dil-LDL）摄取情况。通过网络药理学方法对丹知青娥方治疗高脂血症作用机制进行预测并结合分子对接进行初步验证。建立高脂血症大鼠模型，检测血清总胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）、天冬氨酸氨基转移酶（AST）和丙氨酸氨基转移酶（ALT）水平；蛋白免疫印迹法（Western blot）测定丹知青娥方对高脂血症大鼠肝脏前蛋白转化酶枯草溶菌素9（PCSK9）和低密度脂蛋白受体（LDLR）和蛋白表达情况。[结果] 细胞实验表明，100 μg/mL丹知青娥方显著增强HepG2细胞Dil-LDL摄取能力（P<0.01）。筛选出丹知青娥方药物有效活性成分相应靶点：杜仲2 543个，丹参4 376个，知母1 396个，补骨脂3 217个。高脂血症相关基因靶点2 061个。丹知青娥方与高脂血症共同作用基因靶点324个。基因本体论（GO）分析涉及胆固醇代谢过程，京都基因与基因组百科全书（KEGG）主要与脂质和动脉粥样硬化通路有关，均收录了LDLR基因。分子对接表明PCSK9与核心活性靶点具有良好的对接活性。体内实验表明，10 g/kg丹知青娥方能有效降低高脂血症大鼠的LDL-C、TC、TG、AST、ALT水平以及PCK9蛋白表达，显著上调LDLR蛋白表达（P<0.01）。[结论] 丹知青娥方可能通过调节PCSK9/LDLR信号通路发挥降脂作用。 |
| 关键词: 丹知青娥方 高脂血症 前蛋白转化酶枯草溶菌素9 作用机制 |
| DOI：10.11656/j.issn.1672-1519.2026.07.13 |
| 分类号:R285.5 |
| 基金项目:天津市教委科研计划一般项目（2022KJ182）。 |
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| Research on the lipid-lowering mechanism through the PCSK9/LDLR pathway by Danzhi Qing’e Formula |
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SONG Junli1, SHI Huihui1, CHEN Guangxia2, HUANG Yuxing3, MAO Haoping1, MA Shangwei3
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1.Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China;2.First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300381, China;3.The Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300250, China
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| Abstract: |
| [Objective] To explore the lipid-lowering mechanism of Danzhi Qing’e Formula（DZQEF）based on network pharmacology， molecular docking and experimental studies. [Methods] The uptake of Dil-LDL by HepG2 cells treated with Danzhi Qing’e Formula was detected. The mechanism of Danzhi Qing’e Formula in treating hyperlipidemia was predicted by network pharmacology and preliminarily verified by molecular docking. A hyperlipidemia rat model was established to detect the levels of serum total cholesterol （TC），triglyceride（TG），low-density lipoprotein cholesterol（LDL-C），high-density lipoprotein cholesterol（HDL-C），aspartate aminotransferase（AST）and alanine aminotransferase（ALT）. The protein expressions of proprotein convertase subtilisin/kexin type 9 （PCSK9）and low-density lipoprotein receptor （LDLR）in the liver of hyperlipidemia rats were determined by Western blot. [Results] Cell experiments showed that 100 μg/mL Danzhi Qing’e Formula significantly enhanced the Dil-LDL uptake ability of HepG2 cells（P< 0.01）. A total of 2 543，4 376，1 396 and 3 217 targets were screened out for the effective active components of Eucommia ulmoides， Salvia miltiorrhiza，Anemarrhena asphodeloides and Psoralea corylifolia，respectively. There were 2 061 targets related to hyperlipidemia. There were 324 common targets between Danzhi Qing’e Formula and hyperlipidemia. Gene Ontology（GO）analysis was involved in the cholesterol metabolism process，and Kyoto Encyclopedia of Genes and Genomes（KEGG）was mainly related to lipid and atherosclerosis pathways，both of which included the LDLR gene. Molecular docking indicated that PCSK9 had good docking activity with the core active targets. In vivo experiments showed that 10 g/kg Danzhi Qing’e Formula could effectively reduce LDL-C， TC，TG，AST and ALT levels of hyperlipidemia rats，as well as the protein expressions of PCSK9，and significantly up-regulate the protein expressions of LDLR（P<0.01）. [Conclusion] Danzhi Qing’e Formula may exert lipid-lowering effects by regulating the PCSK9/LDLR signaling pathway. |
| Key words: Danzhi Qing’e Formula dislipidemia proprotein convertase subtilisinkexin type 9 mechanism |
