The Aqueous Extract of Hemerocallis citrina Baroni Improves the Lactation-Promoting Effect in Bovine Mammary Epithelial Cells through the PI3K-AKT Signaling Pathway
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals and Reagents
2.2. Preparation of HAE
2.3. Components in HAE by UHPLC-Q-Exactive Orbitrap-MS Analysis
2.4. Experiments Performed with Animals
2.4.1. Experimental Design and Treatments
2.4.2. The Weight Gain per Litter and the Organ/Tissue Index of Lactating Rats
2.4.3. Detection of Serum Hormone Levels
2.4.4. Histopathological Analysis
2.5. Cell Experimentation
2.5.1. Cell Cultures
2.5.2. Cell Proliferation Assay
2.5.3. Determination of Milk Protein, Lactose, and Milk Fat Content by ELISA
2.6. Network Pharmacology Analysis
2.6.1. Screening of HAE Flavonoid Targets and Lactation-Related Targets
2.6.2. Protein–Protein Interaction (PPI) Network Construction
2.6.3. Core Target Functional Enrichment Analysis
2.7. Molecular Docking
2.8. Transcriptomic Analysis
2.9. DEG Validation by RT-qPCR
2.10. Statistical Analysis
3. Results
3.1. HAE Up-Regulated Lactation Performance in Lactating Rats
3.2. HAE Increased Milk Protein, Lactose, and Milk Fat Contents in BMECs
3.3. RNA-seq Revealed the Regulatory Mechanism of HAE on the Lactation-Promoting Effect of BMECs
3.4. Identification of the Compounds in HAE Based on UHPLC-Q-Exactive Orbitrap-MS
3.5. Network Pharmacology Combined with RNA-seq Revealed HAE Flavonoid-Promoting Effects through Potential Core Targets
3.6. Molecular Docking of HQ and Nine Core Targets
3.7. HQ Up-Regulated Lactation-Promoting Function in BMECs
3.8. Effect of HQ on the Lactation-Related Pathways via DEG Enrichment Analysis
3.9. Analysis of Common DEGs of HAE and HQ
3.10. Effect of HAE and HQ on PI3K-Akt Pathway via RT-qPCR Assays
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Compound Name | Formula | Molecular Weight | CAS | m/z | Ionization Model | Peak Area |
---|---|---|---|---|---|---|
Hexamethylquercetagetin | C21H22O8 | 402.4 | 1251-84-9 | 401.13 | [M − H]− | 421,194,790.80 |
4′,5,6,7,8-Pentahydroxy-3’-methoxyflavone | C16H12O8 | 332.26 | 181020-34-8 | 333.07 | [M + H]+ | 64,114,119.42 |
Rutin | C27H30O16 | 610.5 | 153-18-4 | 611.16 | [M + H]+ | 60,833,174.93 |
Myricetin 3-robinobioside | C27H30O17 | 626.5 | 145544-43-0 | 627.15 | [M + H]+ | 59,991,262.61 |
Licoisoflavone A | C20H18O6 | 354.4 | 66056-19-7 | 353.11 | [M − H]− | 59,916,514.78 |
Myricetin 3-galactoside | C21H20O13 | 480.4 | 15648-86-9 | 481.10 | [M + H]+ | 35,915,744.05 |
Naringenin | C15H12O5 | 272.25 | 480-41-1 | 271.06 | [M − H]− | 18,411,397.14 |
Quercetin | C15H10O7 | 302.23 | 117-39-5 | 301.03 | [M − H]− | 15,911,669.73 |
Kaempferol | C15H10O6 | 286.24 | 520-18-3 | 287.05 | [M + H]+ | 14,296,488.93 |
7-Hydroxy-2-methylisoflavone | C16H12O3 | 252.26 | 2859-88-3 | 253.08 | [M + H]+ | 14,280,814.31 |
Petunidin 3-galactoside | C22H23O12+ | 479.4 | 28500-02-9 | 479.12 | [M + H]+ | 14,378,042.42 |
Daidzin | C21H20O9 | 416.4 | 552-66-9 | 415.11 | [M − H]− | 7,576,634.70 |
Lutein | C40H56O2 | 568.9 | 127-40-2 | 568.43 | [M + H]+ | 10,175,254.17 |
Malvidin 3-(6-acetylglucoside) | C25H27O13+ | 535.5 | 101072-66-6 | 536.16 | [M + H]+ | 10,822,103.81 |
Orientin | C21H20O11 | 448.4 | 28608-75-5 | 449.11 | [M + H]+ | 5,787,667.10 |
Multinoside A | C27H30O16 | 610.5 | 59262-54-3 | 645.12 | [M − H]− | 6,037,243.24 |
Isorhamnetin | C16H12O7 | 316.26 | 480-19-3 | 317.06 | [M + H]+ | 4,581,085.32 |
Myricetin 3-arabinoside | C20H18O12 | 450.3 | 132679-85-7 | 451.09 | [M + H]+ | 4,635,990.00 |
Citroside A | C19H30O8 | 386.4 | 120330-44-1 | 387.20 | [M + H]+ | 3,243,535.37 |
4′,8-Dimethylgossypetin 3-glucoside | C23H24O13 | 508.4 | 90456-57-8 | 509.13 | [M + H]+ | 3,126,823.92 |
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Chen, J.; Pan, Z.; Li, Q.; Wu, Y.; Li, X.; Wang, X.; Hao, D.; Peng, X.; Pan, L.; Li, W.; et al. The Aqueous Extract of Hemerocallis citrina Baroni Improves the Lactation-Promoting Effect in Bovine Mammary Epithelial Cells through the PI3K-AKT Signaling Pathway. Foods 2024, 13, 2813. https://doi.org/10.3390/foods13172813
Chen J, Pan Z, Li Q, Wu Y, Li X, Wang X, Hao D, Peng X, Pan L, Li W, et al. The Aqueous Extract of Hemerocallis citrina Baroni Improves the Lactation-Promoting Effect in Bovine Mammary Epithelial Cells through the PI3K-AKT Signaling Pathway. Foods. 2024; 13(17):2813. https://doi.org/10.3390/foods13172813
Chicago/Turabian StyleChen, Jiaxu, Zhaoping Pan, Qili Li, Yanyang Wu, Xiaopeng Li, Xue Wang, Dandan Hao, Xiaoyu Peng, Lina Pan, Wei Li, and et al. 2024. "The Aqueous Extract of Hemerocallis citrina Baroni Improves the Lactation-Promoting Effect in Bovine Mammary Epithelial Cells through the PI3K-AKT Signaling Pathway" Foods 13, no. 17: 2813. https://doi.org/10.3390/foods13172813