Scientia Agricultura Sinica ›› 2024, Vol. 57 ›› Issue (5): 1010-1022.doi: 10.3864/j.issn.0578-1752.2024.05.015

• ANIMAL SCIENCE·VETERINARY SCIENCE • Previous Articles    

The Potential and Mechanisms of Apigenin to Relieve Heat Stress and Hypoxia in Dairy Cows Based on Network Pharmacology and Molecular Docking

LIU ZhuoLin(), LIU HongYun()   

  1. College of Animal Science, Zhejiang University, The Institute of Dairy Science, Hangzhou 310058
  • Received:2023-05-12 Accepted:2023-07-27 Online:2024-03-06 Published:2024-03-06
  • Contact: LIU HongYun

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Abstract:

【Objective】 The study aimed to predict the mechanisms of apigenin in relieving heat stress and hypoxia stress in dairy cows by using network pharmacology and molecular docking so as to provide a reference for the application of apigenin.【Method】 Firstly, the targets related to apigenin, heat stress, and hypoxia stress were obtained from TCMSP and GeneCards databases, and apigenin targets in relieving dual stress were obtained by Venn intersection. The protein-protein interaction (PPI) network was constructed using the STRING database, and the core targets were identified by analyzing the network node centrality using the CytoNCA plugin in Cytoscape 3.9.1 software. Autodock vina program was used to perform molecular docking of apigenin with the identified targets. Perform gene ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis on the top 30 ranked targets based on degree centrality using Functional Annotation, the analysis tool of David database. Additionally, Molecular complex detection analysis (MCODE) of the above targets yielded the core gene clusters, then GO and KEGG analysis were performed. Apigenin-dual stress targets-signaling pathway network model were constructed through the KEGG pathway enrichment results and core target information. Finally, apigenin was docked to the core targets using the Autodock vina program, and the results were visualized using the Pymol software and the PLIP website.【Result】 The TCMSP database retrieved 68 targets of apigenin, 6 661 hypoxia-related genes and 9 046 heat stress-related genes were obtained in GeneCards database, and 56 apigenin targets in relieving dual stress were obtained. The number of nodes in the target PPI network was 56, the number of edges was 436, the average node degree value was 15.6, the mean local clustering coefficient was 0.728, and PPI enrichment P value was less than 1.0×10-16. The core targets were AKT1 (RAC-alpha serine/ threonine-protein kinase), TP53 (Cellular tumor antigen p53), TNF (Tumor necrosis factor), CASP3 (Caspase-3), INS (Insulin), BCL2L1 (Bcl-2-like protein 1), VEGFA (Vascular endothelial growth factor A), HIF1A (Hypoxia-inducible factor 1-alpha), PTGS2 (Prostaglandin G/H synthase 2), and SREPINE1 (Plasminogen activator inhibitor 1). The molecular docking results showed that apigenin could be stably bound to the above core targets. A total of 54 Biological Processes (BP), 6 Molecular Functions (MF), and 11 Cellular Components (CC) were obtained by GO functional enrichment analysis (P<0.01), and KEGG were enriched to 98 signaling pathways. MCODE analysis showed two major gene clusters. 【Conclusion】 Apigenin mainly regulated cell proliferation, apoptosis, oxygen sensing and inflammation through PI3K-AKT, p53, HIF-1, NF-kappa B and other pathways, to further alleviate heat stress and hypoxia stress. The study provided a theoretical reference for the application of apigenin and apigenin-rich plants in dairy cow production.

Key words: apigenin, dairy cow, heat stress, hypoxia, network pharmacology, molecular docking

Fig. 1

Venn diagram of apigenin targets in relieving dual stress"

Table 1

Information on the core targets of apigenin in relieving dual stress"

序号 Serial number 基因 Gene Uniport ID 名称 Name
1 AKT1 Q01314 RAC-α 丝氨酸/苏氨酸蛋白激酶 RAC-alpha serine/threonine-protein kinase
2 TP53 P67939 细胞肿瘤抗原p53 Cellular tumor antigen p53
3 TNF Q06599 肿瘤坏死因子 Tumor necrosis factor
4 CASP3 Q08DY9 细胞凋亡蛋白酶3 Caspase-3
5 INS P01317 胰岛素 Insulin
6 BCL2L1 Q05KJ0 BCL-2样蛋白1 BCL-2-like protein 1
7 VEGFA P15691 血管内皮生长因子A Vascular endothelial growth factor A
8 HIF1A Q9XTA5 缺氧诱导因子 1-α Hypoxia-inducible factor 1-alpha
9 PTGS2 O62698 前列腺素 G/H 合酶 2 Prostaglandin G/H synthase 2
10 SERPINE1 P13909 纤溶酶原激活物抑制剂 1 Plasminogen activator inhibitor 1

Fig. 2

Network diagram of apigenin targets in relieving dual stress"

Fig. 3

Ranking of TOP 30 degree values for apigenin targets in relieving dual stress"

Fig. 4

Results of GO functional enrichment analysis"

Fig. 5

Results of KEGG enrichment analysis"

Fig. 6

Target protein localization rendering in KEGG enriched pathways"

Fig. 7

Top two gene clusters in MCODE analysis"

Table 2

GO functional analysis of the key gene clusters"

基因簇 Gene Cluster GO条目 GO Term 描述 Description Log10 (P)
MCODE1 GO:0006915 凋亡进程Apoptotic process -6.56
GO:1902004 β-淀粉样蛋白形成的正向调控Positive regulation of amyloid-beta formation -6.45
GO:0005125 细胞因子活性Cytokine activity -5.98
MCODE2 GO:0008353 RNA聚合酶II羧基末端结构域激酶活性
RNA polymerase II CTD heptapeptide repeat kinase activity		 -2.45
GO:0010971 有丝分裂细胞周期G2/M转变的正向调控
Positive regulation of G2/M transition of mitotic cell cycle		 -2.20
GO:0004693 细胞周期蛋白依赖性蛋白丝氨酸/苏氨酸激酶活性
Cyclin-dependent protein serine/threonine kinase activity		 -2.17

Fig. 8

Apigenin-dual stress targets-signaling pathway network model"

Table 3

The binding ability of apigenin to the core targets"

配体
Ligand		 受体
Receptor		 结合自由能
Free energy of banding (kcal·mol-1)
芹菜素
Apigenin		 AKT1 -7.9
TP53 -7.2
TNF -6.1
CASP3 -6.8
INS -7.2
BCL2L1 -7.3
VEGFA -5.9
HIF1A -7.5
PTGS2 -7.6
SERPINE1 -7.2

Fig. 9

Molecular docking results of apigenin to the core targets"

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