槲皮素通过抑制MAPK信号通路改善心力衰竭

doi:10.12122/j.issn.1673-4254.2025.01.22

摘要/Abstract

摘要：

目的基于网络药理学探讨槲皮素治疗心力衰竭的作用机制。方法通过中药系统药理学数据库和Swiss ADME数据库检索槲皮素的作用靶点。利用Genecards、OMIM数据库检索心力衰竭相关靶点。使用DAVID数据库获取交集靶点的GO分析和KEGG信号通路富集分析。交集靶点使用STRING数据库和Cytoscape软件进行蛋白互作分析，并筛选核心基因。最后使用PyMOL与AutoDock Tools软件完成核心靶点与槲皮素的分子对接。以异丙肾上腺素诱导H9C2心肌细胞构建心力衰竭模型，通过细胞实验对筛选出的MAPK信号通路进行验证。结果经筛选共得到60个交集靶点。富集结果显示，槲皮素可能通过MAPK信号通路抑制心力衰竭。核心基因筛选出的AMPK3、BCL-2等可能是槲皮素改善心力衰竭的关键基因。细胞实验表明，不同浓度的槲皮素组呈剂量依赖性地减少异丙肾上腺素诱导心肌细胞的凋亡（P<0.01）。分子生物学实验证实，槲皮素能够降低异丙肾上腺素诱导心肌细胞的Bax/Bcl-2、caspase-3和ERK、p38表达水平（P<0.05）。结论槲皮素可能通过抑制MAPK信号通路抑制心肌细胞凋亡改善心力衰竭。

关键词: 槲皮素, 心力衰竭, 异丙肾上腺素, 丝裂原激活的蛋白激酶, 网络药理学

Abstract:

Objective To explore the mechanism that mediate the therapeutic effect of quercetin on heart failure. Methods We searched the TCMSP and Swiss ADME databases for the therapeutic targets of quercetin and retrieved heart failure targets from the Genecards and OMIM databases. The intersecting targets were analyzed with GO and KEGG pathway analysis using DAVID database, and the key genes were identified via PPI analysis. Molecular docking between the core targets and quercetin was performed using PyMOL and AutoDock Tools. In a heart failure model established in H9C2 cardiomyocytes by treatment with isoproterenol, the effect of quercetin on the expressions of the MAPK signaling pathway was tested. Results A total of 60 intersecting targets were identified. Enrichment analysis revealed that quercetin may inhibit heart failure through the MAPK signaling pathway. The core genes, including AMPK3 and BCL-2, were identified as potential key regulators in quercetin-mediated improvement of heart failure. Cellular experiments demonstrated that quercetin significantly reduced isoproterenol-induced apoptosis of cardiomyocytes in a dose-dependent manner and obviously decreased the Bax/Bcl-2 ratio and the expression levels of caspase-3, ERK and p38 in the cells. Conclusion Quercetin improves heart failure possibly by inhibiting cardiomyocyte apoptosis through the MAPK signaling pathway.

Key words: quercetin, heart failure, isoproterenol, MAPK, network pharmacology

龙秀鹏, 陶顺, 阳绅, 李素云, 饶利兵, 李莉, 张哲. 槲皮素通过抑制MAPK信号通路改善心力衰竭[J]. 南方医科大学学报, 2025, 45(1): 187-196.

Xiupeng LONG, Shun TAO, Shen YANG, Suyun LI, Libing RAO, Li LI, Zhe ZHANG. Quercetin improves heart failure by inhibiting cardiomyocyte apoptosis via suppressing the MAPK signaling pathway[J]. Journal of Southern Medical University, 2025, 45(1): 187-196.

图/表 10

表1 引物序列

Tab.1 Primer sequence for qRT-PCR

Genes	Primer sequence
Bcl-2	F: 5'-ATCTCCCTGTTGACGCTCT-3′
	R: 5'-CATCTTCTCCTTCCAGCCT-3′
Bax	F: 5'-AGCCACAAAGATGGTCACT-3′
	R: 5'-GGAGATGAACTGGATAGCAA-3′
Caspase-3	F: 5'-TGTTTCCCTGAGGTTTGCTG-3′
	R: 5'-TGCTATTGTGAGGCGGTTGT-3′
ERK	F: 5'-CGGAACTTGCAATCCTCAGT-3′
	R: 5'-TCGTGTGGGTCCTGAATTGG-3′
p38	F: 5'-CTGCGAGGGCTGAAGTAT-3′
	R: 5'-TCCTCTTATCCGAGTCCAA-3′
GAPDH	F: 5'-GAAGGTGAAGGTCGGAGTC-3'
	R: 5'-GAAGATGGTGATGGGATTTC-3'

图1 槲皮素与心衰的交集靶点

Fig.1 Intersection targets of quercetin and heart failure. Que: Quercetin; HF: Heart failure.

图2 槲皮素与心力衰竭的核心基因

Fig.2 Core genes of quercetin for treatment of heart failure.

图4 核心靶点-槲皮素分子对接结果

Fig.4 Molecular docking results of the core targets with quercetin.

图5 不同浓度槲皮素对H9C2心肌细胞存活率的影响

Fig.5 Effects of different concentrations of quercetin on survival rate of H9C2 cardiomyocytes. **P<0.01 vs 0 μmol/L group (n=5).

图6 不同浓度槲皮素对ISO诱导心肌细胞损伤的影响

Fig.6 Effects of different concentrations of quercetin(Que) on isoproterenol (ISO)-induced injury in mouse cardiomyocytes. A: Effects of different concentrations of quercetin on the survival rate of myocardial cells induced by ISO. B: Hoechst staining for detecting apoptosis of the injured cardiomyocytes (Original magnification:×200). ##P<0.01 vs Control group, *P<0.05, **P<0.01 vs ISO group (n=3).

图8 不同浓度槲皮素对各组心肌细胞中Bax、Bcl-2、caspase-3 mRNA表达的影响

Fig.8 Effects of different concentrations of quercetin on mRNA expressions of Bcl-2 (A), Bax (B), Bax/Bcl-2 ratio (C) and caspase-3 (D) in the cardiomyocytes in each group. ##P<0.01 vs Control group, *P<0.05, **P<0.01 vs ISO group (n=3).

图9 不同浓度槲皮素对各组心肌细胞凋亡相关蛋白表达的影响

Fig.9 Effects of different concentrations of quercetin on the expression of apoptosis-related proteins in the cardiomyocytes in each group. A: Western blotting of c-caspase-3, t-caspase-3, Bcl-2 and Bax proteins. B: Ratio of cleaved-caspase-3/caspase-3 protein. C: Ratio of Bax/Bcl-2 protein. ##P<0.01 vs Control group, **P<0.01 vs ISO group (n=3).

图10 不同浓度槲皮素对各组心肌细胞ERK/p38表达的影响

Fig.10 Effects of different concentrations of quercetin on ERK/p38 expression in the cardiomyocytes in each group. A, B: Effects of quercetin on expressions of ERK and p38 mRNA in the cells. C: Western blotting of p-ERK, ERK, p-p38 and p38 proteins. D, E: Expression of p-ERK/ERK and p-p38/p38 protein in the cells. ##P<0.01 vs Control group, *P<0.05, **P<0.01, ***P<0.001 vs ISO group (n=3).

图11 不同浓度槲皮素对ERK/p38通路抑制剂介导各组心肌细胞凋亡相关蛋白的表达影响

Fig.11 Effects of different concentrations of quercetin on expressions of apoptosis-related proteins in cardiomyocytes treated with ERK/p38 pathway inhibitors. A: Effects of different concentrations of quercetin on the expression of Bax, Bcl-2 and caspase-3 in the cells after adding ERK inhibitor (TBHQ) and p38 inhibitor (SB 203580). B: Ratio of Bax/Bcl-2 protein expression. C: Ratio of cleaved-caspase-3/caspase-3 protein expression. ##P<0.01 vs Control group, **P<0.01 vs ISO group, △△P<0.01 vs ISO+Que group (n=3).

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