清心牛黄丸通过改善脂质代谢紊乱缓解小鼠非酒精性脂肪性肝病

doi:10.12122/j.issn.1673-4254.2025.09.04

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (9): 1840-1849.doi: 10.12122/j.issn.1673-4254.2025.09.04

• • 上一篇

清心牛黄丸通过改善脂质代谢紊乱缓解小鼠非酒精性脂肪性肝病

骆碧云¹^,²(), 易欣¹, 蔡怡静¹, 张世卿³, 王鹏², 李彤², 翁建霖⁴^,⁵, 周平正¹()

^1.南方医科大学药学院，广东广州 510515
^2.南方医科大学第五附属医院药学部，广东广州 510925
^3.暨南大学药学院，广东广州 510632
^4.香港教育大学科学与环境学系，香港 999077
^5.香港浸会大学金卫医疗神经再生研究中心，香港 999077

收稿日期:2025-02-27 出版日期:2025-09-20 发布日期:2025-09-28
通讯作者: 周平正 E-mail:wfyyjk@163.com;pzzhou@smu.edu.cn
作者简介:骆碧云，在读硕士研究生，E-mail: wfyyjk@163.com
基金资助:
国家自然科学基金与香港研究资助局合作研究项目(8231101099);香港研究资助局研究配对补助金计划(RMGS2021-10-13);香港研究资助局研究配对补助金计划(RMGS2023-5-012)

Ching Shum Pills alleviates non-alcoholic fatty liver disease in mice by ameliorating lipid metabolism disorders

Biyun LUO¹^,²(), Xin YI¹, Yijing CAI¹, Shiqing ZHANG³, Peng WANG², Tong LI², Ken Kin Lam YUNG⁴^,⁵, Pingzheng ZHOU¹()

^1.School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China
^2.Department of Pharmacy, Fifth Affiliated Hospital of Southern Medical University, Guangzhou 510925, China
^3.College of Pharmacy, Jinan University, Guangzhou 510632, China
^4.Department of Science and Environmental Studies, the Education University of Hong Kong, Hong Kong 999077, China
^5.Golden Meditech Centre for Neuro-Regeneration Sciences, Hong Kong Baptist University, Hong Kong 999077, China

Received:2025-02-27 Online:2025-09-20 Published:2025-09-28
Contact: Pingzheng ZHOU E-mail:wfyyjk@163.com;pzzhou@smu.edu.cn

摘要/Abstract

摘要：

目的探讨清心牛黄丸对非酒精性脂肪性肝病（NAFLD）的干预作用及潜在机制。方法建立NAFLD小鼠模型，采用清心牛黄丸进行干预治疗，并检测血糖、血脂及肝功能等血清生化指标以评估其药效。结合网络药理学方法初步预测清心牛黄丸的关键活性成分及潜在作用靶点，相关化学成分来源于TCMSP和HERB数据库，基于SwissTargetPrediction平台预测靶点，通过GeneCards、OMIM和DisGeNET数据库筛选获得疾病靶点。构建蛋白－蛋白互作（PPI）网络，进行GO和KEGG功能富集分析。采用AutoDock Vina进行关键成分与靶点的分子对接，根据对接结果，通过实时荧光定量PCR（RT-qPCR）验证核心靶点的表达变化，探讨清心牛黄丸的作用机制。结果与高脂对照组相比，低、高剂量清心牛黄丸抑制小鼠体质量增长（P<0.05），降低血清TG水平（P<0.01）；高剂量清心牛黄丸降低小鼠血清ALT水平（P<0.05），抑制脂肪积累（P<0.001）。网络药理学预测发现，清心牛黄丸治疗NAFLD的核心成分为槲皮素和亚麻酸单酰甘油。分子对接结果显示，这两种核心成分能与TNF、AKT1、IL6、TP53、ALB等5个核心靶蛋白稳定结合，支持其在治疗NAFLD中起关键作用。qPCR实验结果显示，高脂饮食抑制上述基因的表达，而清心牛黄丸干预可明显恢复其表达水平。结论清心牛黄丸可通过多靶点、多通路的调控机制改善NAFLD小鼠的脂质代谢紊乱，减轻肝脏脂肪沉积，并在一定程度上改善肝功能。其中，槲皮素和亚麻酸单酰甘油等关键活性成分可能通过靶向TP53等核心靶点，参与脂肪酸氧化等代谢过程从而发挥治疗作用。

关键词: 非酒精性脂肪性肝病, 清心牛黄丸, 槲皮素, 脂质代谢, 肝功能

Abstract:

Objective To investigate the effect of Ching Shum Pills (CSP) for alleviating non-alcoholic fatty liver disease (NAFLD) and the underlying mechanism. Methods In a mouse model of NAFLD, the therapeutic effect of CSP was evaluated by measuring serum glucose, lipid profiles (TC, TG, LDL-C, HDL-C), and hepatic function markers. Network pharmacology was employed to identify active compounds in CSP and their targets using TCMSP, HERB, SwissTargetPrediction, GeneCards, OMIM, and DisGeNET. Protein-protein interaction (PPI) networks, Gene Ontology (GO), and KEGG pathway analyses were conducted. Molecular docking (AutoDock Vina) was used to assess the compound-target binding affinities. Quantitative real-time PCR (qRT-PCR) was used to validate the mRNA expressions of the core genes in the liver tissue of the mouse models. Results In the mouse model of NAFLD, treatment with CSP significantly reduced body weight gain and serum TG levels of the mice, and high-dose CSP treatment resulted in obvious reduction of ALT levels and hepatic fat accumulation. Network pharmacology analysis identified quercetin and 2-monolinolenin as the key bioactives in CSP, which target TNF, AKT1, IL6, TP53, and ALB. Docking simulations suggested strong binding between the two core compounds and their target proteins. The results of qRT-PCR showed that high-fat diet induced significant downregulation of Tp53, Cpt1, and Ppara expressions in mice, which was effectively reversed by CSP treatment. Conclusion CSP can improve lipid metabolism disorders in NAFLD mice through a regulatory mechanism involving multiple targets and pathways to reduce liver fat accumulation and protect liver function. The key components in CSP such as quercetin and linolenic acid monoacylglycerol may participate in the regulation of such metabolic processes as fatty acid oxidation by targeting TP53.

Key words: non-alcoholic fatty liver disease, Ching Shum Pills, quercetin, lipid metabolism, liver function

骆碧云, 易欣, 蔡怡静, 张世卿, 王鹏, 李彤, 翁建霖, 周平正. 清心牛黄丸通过改善脂质代谢紊乱缓解小鼠非酒精性脂肪性肝病[J]. 南方医科大学学报, 2025, 45(9): 1840-1849.

Biyun LUO, Xin YI, Yijing CAI, Shiqing ZHANG, Peng WANG, Tong LI, Ken Kin Lam YUNG, Pingzheng ZHOU. Ching Shum Pills alleviates non-alcoholic fatty liver disease in mice by ameliorating lipid metabolism disorders[J]. Journal of Southern Medical University, 2025, 45(9): 1840-1849.

图/表 8

表1 小鼠qPCR引物序列

Tab.1 Primer sequence for RT-qPCR in this study

Gene	Forward/Reverse primer	Primer sequences (5'-3')
β-actin	Forward	AAGTCCCTCACCCTCCCAAAAG
	Reverse	AAGCAATGCTGTCACCTTCCC
Tp53	Forward	GTCACAGCACATGACGGAGG
	Reverse	TCTTCCAGATACTCGGGATAC
Cpt1	Forward	TCCACCCTGAGGCATCTATT
	Reverse	ATGACCTCCTGGCATTCTCC
Ppara	Forward	CGGGAAAGACCAGCAACAAC
	Reverse	ATAGCAGCCACAAACAGGGA

图1 CSP缓解高脂饮食诱导的小鼠体质量增加

Fig.1 CSP mitigates high-fat diet-induced non-alcoholic fatty liver disease (NAFLD) in mice. A: Schematic diagram of NAFLD modeling and CSP treatment in mice. B: Body weight changes in mice during the experiment. C: Percentage of epididymal fat pad weight in body weight. D: Average food intake during the experiment. E: Weight of liver after indicated treatment. F: Fasting plasma glucose. G: Postprandial plasma glucose. ***P<0.001; #P <0.05.

图2 各组小鼠血清生化指标

Fig.2 Serum biochemical parameters of the mice in each group. A: Triglycerides (TG) in serum. B: Total cholesterol (TC) in serum. C: Low-density lipoprotein cholesterol (LDL-C) in serum. D: High-density lipoprotein cholesterol (LDL-C) in serum. E: Alanine aminotransferase (ALT) in serum. F: Aspartate aminotransferase (AST) in serum. *P<0.05, **P <0.01, ***P<0.001 vs HFD group.

图3 各组小鼠肝脏病理染色情况

Fig.3 Histopathological staining of the liver tissues of the mice in each group. A: HE staining and Oil Red O staining of the liver tissues. B: HE bubble vacuole ratio in each group. C: Quantification of Oil Red O stain coverage in each group. *P<0.001 vs HFD group; ###P<0.001 vs Control group.

图4 清心牛黄丸-非酒精性脂肪性肝病的共同靶点

Fig.4 Common targets of Ching Shum Pill (CSP) and non-alcoholic fatty liver disease (NAFLD). A: Venn diagram of CSP- and NAFLD-related targets. B: Top 30 key targets of CSP active ingredients in NAFLD treatment.

图5 "中药－化学成分－关键靶点－疾病"网络

Fig.5 "Herb-compound-target-disease" network of CSP.

图6 CSP治疗NAFLD的关键靶点富集分析

Fig.6 Key target enrichment analysis of CSP for NAFLD treatment. A: GO enrichment analysis, including biological process (BP), cellular components (CC) and molecular function (MF). B: KEGG enrichment analysis.

图7 分子对接和RT-qPCR检测小鼠肝组织Tp53、Cpt1和Ppara的表达水平

Fig.7 Molecular docking and RT-qPCR analysis of the expression levels of Tp53, Cpt1, and Ppara in mouse liver tissues. A: Docking between AKT1 and monolinolenin. B: Docking between ALB-2 and monolinolenin. C: Docking between IL-6 and monolinolenin. D: Docking between TP53 and monolinolenin. E: Docking between TP53 and quercetin. F: Tp53 mRNA expression. G: Cpt1 mRNA expression. H: Ppara mRNA expression. Data are normalized to β-actin and presented as relative values to the mean of the HFD group. *P<0.05, **P<0.01, ***P<0.001.

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清心牛黄丸通过改善脂质代谢紊乱缓解小鼠非酒精性脂肪性肝病

Ching Shum Pills alleviates non-alcoholic fatty liver disease in mice by ameliorating lipid metabolism disorders

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