Astragaloside IV Alleviates Ammonia-Induced Apoptosis and Oxidative Stress in Bovine Mammary Epithelial Cells
Abstract
:1. Introduction
2. Results
2.1. Effect of Astragaloside IV on Ammonia-Induced Bovine Mammary Epithelial Cell Death
2.2. Effects of Astragaloside IV on mRNA Expressions of Apoptosis-Related Genes Induced by Ammonia in Bovine Mammary Epithelial Cells
2.3. Effects of Ammonia and Astragaloside IV Treatment on p53 Signaling Pathways in Bovine Mammary Epithelial Cells
2.4. Effects of Astragaloside IV on mRNA Expression of Inflammatory Factors (IL-6 and IL-8) Induced by Ammonia in Bovine Mammary Epithelial Cells
2.5. Effects of Astragaloside IV on mRNA Expression of Endoplasmic Reticulum Stress Markers (CHOP and GPR78) Induced by Ammonia in Bovine Mammary Epithelial Cells
2.6. The Cytoprotective Effects of Astragaloside IV against Oxidative Stress are Dependent on the Induction of Nrf2 in Bovine Mammary Epithelial Cells
2.7. Effects of Ammonia and Astragaloside IV Treatment on the AKT and ERK Signaling Pathways in Bovine Mammary Epithelial Cells
2.8. Activation of AKT and ERK Is Required for the Cytoprotective Effects of Astragaloside IV in Bovine Mammary Epithelial Cells
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatment
4.2. Cell Viability Assay
4.3. Detection of Intracellular ROS
4.4. Quantitative Measurement of Apoptosis by AnnexinV-FITC/PI Staining
4.5. RNA Isolation and Quantitative Real-Time PCR (qPCR)
4.6. Transfection and Nrf2 Small RNA Interference
4.7. Western Blot Analysis
4.8. Immunofluorescent Staining
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
ROS | Reactive oxygen species |
CHOP | C/EBP homologous protein |
GRP78 | Glucose-regulated protein 78 |
HO-1 | Hemeoxygenase 1 |
Nrf2 | Nuclear factor erythroid 2-related factor 2 |
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Primer Name | Primer Sequence (5′-3′) | Product Length (bp) |
---|---|---|
BAX | Sense Primer: TGCTTCAGGGTTTCATCC Anti-sense Primer: CTTCAGACACTCGCTCAG | 116 |
BCL-2 | Sense Primer: TTCTCCTGGCTGTCTCTG Anti-sense Primer: CTGCTTCTTGAATCTTCTGC | 135 |
caspase 3 | Sense Primer: GTTCATCCAGGCTCTTTGTG Anti-sense Primer: AAGGACTCATATTCTATTGCTACC | 108 |
p53 | Sense Primer: GAAGACCTACCCTGGCAATTAC Anti-sense Primer: AGAACAGCTTGTTAAGGGAAGG | 104 |
GAPDH | Sense Primer: GTTCAACGGCACAGTCAAG Anti-sense Primer: TACTCAGCACCAGCATCAC | 117 |
Nrf2 | Sense Primer: CCAGCACAACACATACCATCAG Anti-sense Primer: CGTAGCCGAAGAAACCTCATTG | 156 |
CHOP | Sense Primer: TGAACGACTCAAACAGGAAATC Anti-sense Primer: ACGCTAAGACCCTTTTCTATCG | 248 |
GPR78 | Sense Primer: GACCCTGACTCGGGCTAAAT Anti-sense Primer: TGGACAGCGGCACCATATG | 243 |
HO-1 | Sense Primer: GGCAGCAAGGTGCAAGA Anti-sense Primer: GAAGGAAGCCAGCCAAGAG | 221 |
xCT | Sense Primer: GATACAAACGCCCAGATATGC Anti-sense Primer: ATGATGAAGCCAATCCCTGTA | 136 |
IL6 | Sense Primer: ATGCTTCCAATCTGGGTTC Anti-sense Primer: TGAGGATAATCTTTGCGTTC | 269 |
IL8 | Sense Primer: GCTGGCTGTTGCTCTCTTG Anti-sense Primer: GGGTGGAAAGGTGTGGAATG | 126 |
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Wang, F.; Zhao, Y.; Chen, S.; Chen, L.; Sun, L.; Cao, M.; Li, C.; Zhou, X. Astragaloside IV Alleviates Ammonia-Induced Apoptosis and Oxidative Stress in Bovine Mammary Epithelial Cells. Int. J. Mol. Sci. 2019, 20, 600. https://doi.org/10.3390/ijms20030600
Wang F, Zhao Y, Chen S, Chen L, Sun L, Cao M, Li C, Zhou X. Astragaloside IV Alleviates Ammonia-Induced Apoptosis and Oxidative Stress in Bovine Mammary Epithelial Cells. International Journal of Molecular Sciences. 2019; 20(3):600. https://doi.org/10.3390/ijms20030600
Chicago/Turabian StyleWang, Fengge, Yun Zhao, Shuxiong Chen, Lu Chen, Liting Sun, Maosheng Cao, Chunjin Li, and Xu Zhou. 2019. "Astragaloside IV Alleviates Ammonia-Induced Apoptosis and Oxidative Stress in Bovine Mammary Epithelial Cells" International Journal of Molecular Sciences 20, no. 3: 600. https://doi.org/10.3390/ijms20030600