Pcsk6 Deficiency Promotes Cardiomyocyte Senescence by Modulating Ddit3-Mediated ER Stress
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animal Experiments
2.2. Cell Culture and Treatment
2.3. SiRNA and Plasmid DNA Transfection
2.4. RNA Sequencing and Analysis
2.5. Immunofluorescent Staining
2.6. Hoechst Staining
2.7. Senescence-Associated β-Galactosidase (SA-β-Gal) Staining
2.8. RNA Isolation and Quantitative Real-Time PCR (qPCR)
2.9. Protein Extraction and Western Blotting
2.10. Measurement of Reactive Oxygen Species (ROS)
2.11. Measurement of Apoptotic Activity
2.12. Statistical Analysis
3. Results
3.1. PCSK6 Protein Expression Is Reduced in the Heart of Aged Mice
3.2. PCSK6 Expression Is Decreased in the Senescent Cardiomyocytes Induced by D-Gal
3.3. Pcsk6 Gene Knockdown Promotes Cardiomyocyte Senescence
3.4. Pcsk6 Gene Knockdown Impairs Cardiomyocyte Function
3.5. PCSK6 Overexpression Reverses Cardiomyocyte Senescence and Dysfunction Induced by D-gal
3.6. Identification of the Ddit3 Gene Related to Protein Processing in the Endoplasmic Reticulum (ER) upon Pcsk6-Knockdown
3.7. PCSK6 Regulates DDIT3 Expression in Tunicamycin (Tm)-Treated Cardiomyocytes
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Zhan, W.; Chen, L.; Liu, H.; Long, C.; Liu, J.; Ding, S.; Wu, Q.; Chen, S. Pcsk6 Deficiency Promotes Cardiomyocyte Senescence by Modulating Ddit3-Mediated ER Stress. Genes 2022, 13, 711. https://doi.org/10.3390/genes13040711
Zhan W, Chen L, Liu H, Long C, Liu J, Ding S, Wu Q, Chen S. Pcsk6 Deficiency Promotes Cardiomyocyte Senescence by Modulating Ddit3-Mediated ER Stress. Genes. 2022; 13(4):711. https://doi.org/10.3390/genes13040711
Chicago/Turabian StyleZhan, Wenxing, Liping Chen, Hongfei Liu, Changkun Long, Jiankun Liu, Shuangjin Ding, Qingyu Wu, and Shenghan Chen. 2022. "Pcsk6 Deficiency Promotes Cardiomyocyte Senescence by Modulating Ddit3-Mediated ER Stress" Genes 13, no. 4: 711. https://doi.org/10.3390/genes13040711