金银花提取物对阿霉素诱导的小鼠心肌损伤的保护作用及其机制

doi:10.12122/j.issn.1673-4254.2025.12.01

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (12): 2527-2540.doi: 10.12122/j.issn.1673-4254.2025.12.01

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金银花提取物对阿霉素诱导的小鼠心肌损伤的保护作用及其机制

夏士程¹^,³(), 韦慧芳¹^,³, 洪维灿¹^,³, 张钰明²^,³, 尹菲玚¹^,³, 张贻欣¹^,³, 张淋淋²^,³, 高琴²^,³(), 叶红伟²^,³()

^1.蚌埠医科大学，临床医学院，安徽蚌埠 233030
^2.蚌埠医科大学，生理学教研室，安徽蚌埠 233030
^3.蚌埠医科大学，心脑血管疾病基础与临床重点实验室，安徽蚌埠 233030

收稿日期:2025-03-04 接受日期:2025-07-15 出版日期:2025-12-20 发布日期:2025-12-22
通讯作者: 高琴,叶红伟 E-mail:12210110342@stu.bbmc.edu.cn;bbmcgq@126.com;yehongwei223@163.com
作者简介:夏士程，本科，E-mail：12210110342@stu.bbmc.edu.cn

Protective effect of Lonicerae Japonicae Flos extract against doxorubicin-induced myocardial injury in mice and the possible mechanisms

Shicheng XIA¹^,³(), Huifang WEI¹^,³, Weican HONG¹^,³, Yuming ZHANG²^,³, Feiyang YIN¹^,³, Yixin ZHANG¹^,³, Linlin ZHANG²^,³, Qin GAO²^,³(), Hongwei YE²^,³()

^1.Department of Clinical Medicine, Bengbu Medical University, Bengbu 233030, China
^2.Department of Physiology, Bengbu Medical University, Bengbu 233030, China
^3.Key Laboratory of Basic and Clinical Research of Cardiovascular and Cerebrovascular Diseases, Bengbu Medical University, Bengbu 233030, China

Received:2025-03-04 Accepted:2025-07-15 Online:2025-12-20 Published:2025-12-22
Contact: Qin GAO, Hongwei YE E-mail:12210110342@stu.bbmc.edu.cn;bbmcgq@126.com;yehongwei223@163.com
Supported by:
Anhui Provincial Funds for Excellent Scientific Research and Innovation Team(2022AH010083);安徽省高校优秀科研创新团队项目(2022AH010083);512 Talent Program of Bengbu Medical University(by51201102);蚌埠医科大学“512人才计划”(by51201102);Innovation and Entrepreneurship Training Program for College Students(S202410367081);大学生创新创业训练计划项目(S202410367081);Innovation and Entrepreneurship Training Program for College Students(S202410367011);Innovation and Entrepreneurship Training Program for College Students(202410367058);Innovation and Entrepreneurship Training Program for College Students(S202410367057)

摘要/Abstract

摘要：

目的探讨金银花（LJF）对阿霉素（DOX）诱导的心肌损伤的保护作用及其机制。方法通过网络药理学、生物信息学分析与分子对接技术预测核心靶点，并通过动物实验加以验证。动物实验中，检测DOX诱导的心肌损伤及不同剂量LJF提取物治疗后小鼠心功能、心肌酶学、心肌组织形态、炎症因子及相关蛋白表达的变化。结果网络药理学筛选出LJF的10个核心活性成分可与AKT、EGFR、GSK3β良好结合。动物实验结果显示，与假手术组相比，DOX组小鼠心输出量、每搏输出量、左室射血分数及左室短轴缩短率显著降低，血清CK-MB、LDH水平升高，心肌IL-18、IL-1β含量增加；HE染色示心肌结构损伤；心肌组织NLRP3、caspase-1、GSDMD及GSDMD-N蛋白表达上调，EGFR蛋白表达下调，p-AKT、p-GSK3β蛋白水平降低。与DOX组相比，LJF治疗后小鼠心功能明显改善，心肌组织中IL-18、IL-1β水平降低，NLRP3、caspase-1、GSDMD及GSDMD-N蛋白表达下调， EGFR蛋白水平上调，p-AKT、p-GSK3β蛋白磷酸化水平提高。结论金银花可能通过靶向作用于EGFR、AKT、GSK3β调控ErbB信号通路，抑制心肌组织炎症反应与细胞焦亡，从而减轻阿霉素诱导的心肌损伤。

关键词: 网络药理学, 金银花提取物, 阿霉素, 分子对接, 细胞焦亡, ErbB信号转导通路

Abstract:

Objective To evaluate the protective effect of Lonicerae Japonicae Flos (LJF) extract against doxorubicin (DOX)-induced cardiotoxicity (DIC) and explore the possible mechanisms. Methods Network pharmacology, bioinformatics analysis and molecular docking were used to predict the targets of the core components of LJF. In a mouse model of DOX-induced myocardial injury, the protective effects of different doses of LJF extract were evaluated and the underlying mechanisms were explored by detecting the changes in mouse myocardial functions, myocardial enzymes, myocardial pathologies, and the expressions of inflammatory factors and pyroptosis-related proteins. Results The 10 core ingredients of LJF showed strong binding to AKT, EGFR, and GSK3β. In the animal experiment, the DOX-treated mice, compared with the sham-treated mice, had significantly decreased cardiac output, stroke volume, left ventricular ejection fraction, left ventricular fraction shorting, elevated serum levels of CK-MB and LDH, increased myocardial expressions of IL-18 and IL-1β, obvious myocardial damage, increased expression levels of NLRP3, caspase-1, GSDMD and GSDMD-N, and reduced expressions of EGFR, p-AKT and p-GSK3β proteins in the myocardial tissues. LJF treatment obviously improved myocardial function, decreased myocardial expressions of IL-18, IL-1β, NLRP3, caspase-1, GSDMD and GSDMD-N proteins, and increased the expressions EGFR, p-AKT and p-GSK3β proteins in DOX-treated mice. Conclusion LJF extract alleviates DOX-induced myocardial injury in mice possibly by reducing myocardial inflammation and pyroptosis via targeting EGFR, AKT and GSK3β to regulate the ErbB signaling pathway.

Key words: network pharmacology, Lonicerae Japonicae Flos extract, doxorubicin, molecular docking, pyroptosis, ErbB signaling pathway

夏士程, 韦慧芳, 洪维灿, 张钰明, 尹菲玚, 张贻欣, 张淋淋, 高琴, 叶红伟. 金银花提取物对阿霉素诱导的小鼠心肌损伤的保护作用及其机制[J]. 南方医科大学学报, 2025, 45(12): 2527-2540.

Shicheng XIA, Huifang WEI, Weican HONG, Yuming ZHANG, Feiyang YIN, Yixin ZHANG, Linlin ZHANG, Qin GAO, Hongwei YE. Protective effect of Lonicerae Japonicae Flos extract against doxorubicin-induced myocardial injury in mice and the possible mechanisms[J]. Journal of Southern Medical University, 2025, 45(12): 2527-2540.

图/表 12

Fig.1 Flow chart of the study design.

Tab.1 Main active components and the core ingredients in Lonicerae Japonicae Flos （LJF）

Mol ID	Molecule	OB	DL	Core ingredients
MOL000006	luteolin	36.16	0.25	Yes
MOL000098	quercetin	46.43	0.28	Yes
MOL000358	beta-sitosterol	36.91	0.75	No
MOL000422	kaempferol	41.88	0.24	Yes
MOL000449	Stigmasterol	43.83	0.76	No
MOL001494	Mandenol	42	0.19	No
MOL001495	Ethyl linolenate	46.1	0.2	No
MOL002707	phytofluene	43.18	0.5	No
MOL002773	beta-carotene	37.18	0.58	No
MOL002914	eriodyctiol (flavanone)	41.35	0.24	Yes
MOL003006	(-)-(3R,8S,9R,9aS,10aS)-9-ethenyl-8-(beta-D-glucopyranosyloxy)-2,3,9,9a,10, 10a-hexahydro-5-oxo-5H,8H-pyrano[4,3-d]oxazolo[3,2-a]pyridine-3-carboxylic acid_qt	87.47	0.23	Yes
MOL003014	secologanic dibutylacetal_qt	53.65	0.29	Yes
MOL003036	ZINC03978781	43.83	0.76	No
MOL003044	chryseriol	35.85	0.27	Yes
MOL003059	kryptoxanthin	47.25	0.57	No
MOL003062	4,5'-Retro-.beta.,.beta.-Carotene-3,3'-dione, 4',5'-didehydro-	31.22	0.55	No
MOL003095	5-hydroxy-7-methoxy-2-(3,4,5-trimethoxyphenyl)chromone	51.96	0.41	Yes
MOL003101	7-epi-Vogeloside	46.13	0.58	No
MOL003108	caeruloside C	55.64	0.73	No
MOL003111	centauroside_qt	55.79	0.5	Yes
MOL003117	niceracetalides B_qt	61.19	0.19	Yes
MOL003124	XYLOSTOSIDINE	43.17	0.64	No
MOL003128	dinethylsecologanoside	48.46	0.48	No

Fig.2 Target genes of the core ingredients in LJF (A) and GO enrichment analysis (B) and KEGG enrichment analysis (C) of these targets.

Tab.2 Top 10 target genes based on degree values

Gene name	Betweenness unDir	Closeness unDir	Degree unDir
Akt1	3568.885	0.004608	83
Stat3	1519.647	0.004149	66
Pparg	1574.181	0.004032	57
Gsk3b	2449.064	0.004082	55
Egfr	632.9566	0.003906	54
Esr1	1016.3	0.003861	50
Mmp9	1198.398	0.003861	49
Hif1a	515.0458	0.003876	49
Ptgs2	971.437	0.003861	49
Mapk14	546.9512	0.003831	48

Fig.3 Venn diagram of the target genes of LJF, adriamycin-induced myocardial injury and pyroptosis-related targets (A) and the intersection genes network diagram (B).

Fig.4 GO enrichment analysis (A) and KEGG enrichment analysis (B) of the intersection genes and diagram of the ErbB signaling pathway (C).

Tab.3 Binding energy of the core ingredients of LJF with their core target genes

Mol ID	Binding energy (kJ·mol^-1)
Mol ID	EGFR	AKT1	GSK3β
MOL000006	-5.51	-6.98	-5.83
MOL000098	-6.07	-6.96	-5.8
MOL000422	-5.87	-7.41	-6.45
MOL002914	-5.46	-5.36	-5.23
MOL003006	-5.81	-5.92	-5.06
MOL003014	-2.13	-3.02	-2.25
MOL003044	-4.83	-7.2	-5.66
MOL003095	-5	-6.82	-5.49
MOL003111	-6.29	-3.99	-3.37
MOL003117	-5.91	-5.31	-4.8

Fig.5 Molecular docking study of the core ingredients of LJF with their core target genes and their interactions. A: Part of the results of molecular docking results. B: The "drug-component-site of action-type of action-protein" network of EGFR. C: The "drug-component-site of action-type of action-protein" network of GSK3β. D: The "drug-component-site of action-type of action-protein" network of AKT.

Fig. 6 Cardiac echocardiographic findings of the mice and the cardiac function parameters of the mice in each group. A: Cardiac echocardiography of the mice in each group. B: Comparison of cardiac output of the mice among the 5 groups. C: Comparison of the stroke volume of the mice among the 5 groups. D: Comparison of ejection fraction of the mice among the 5 groups. E: Comparison of fraction shorting of the mice among the 5 groups. Data are presented as Mean±SD (n=6). ****P<0.0001 vs Sham group; #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001 vs DOX group.

Fig.7 HE staining of myocardial tissue in each group of mice (Original magnification: ×20).

Fig.8 Serum levels of CK-MB (A) and LDH (B) and the levels of IL-18 (C) and IL-1β (D) in the myocardial tissue of the mice in each group (Mean±SD, n=6). ****P<0.0001 vs Sham group; #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001 vs the DOX group.

Fig.9 Expression levels of NLRP3, caspase-1, GSDMD, GSDMD-N, EGFR, AKT, GSK3β, p-AKT and p-GSK3β in the myocardial tissues detected by Western blotting. A-D: Western blotting for detecting the expression levels of EGFR, AKT, p-AKT, GSK3β and p-GSK3β proteins in each group. E-I: Western blotting for detecting the expression levels of NLRP3, caspase-1, GSDMD and GSDMD-N proteins in each group. ***P<0.001, **P<0.01, *P<0.05 vs Sham group; ##P<0.01, #P<0.05 vs DOX group (Mean±SD, n=3).

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金银花提取物对阿霉素诱导的小鼠心肌损伤的保护作用及其机制

Protective effect of Lonicerae Japonicae Flos extract against doxorubicin-induced myocardial injury in mice and the possible mechanisms

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