MicroRNA-223 Suppresses Human Hepatic Stellate Cell Activation Partly via Regulating the Actin Cytoskeleton and Alleviates Fibrosis in Organoid Models of Liver Injury
Abstract
:1. Introduction
2. Results
2.1. Ectopic miR-223 Expression Suppresses Activation of Human HSCs
2.2. miR-223 Promotes Quiescent Features of Human HSCs
2.3. miR-223 Suppressed Cytoskeletal Activity of Primary Human HSCs
2.4. miR-223 Directly Targets ACTA2 and Inhibits Erk1/2 and Smad2/3 Phosphorylation
2.5. Restoration of α-SMA Inhibits the Antifibrotic Effects of miR-223
2.6. miR-223 Blocks Fibrogenesis Induced by Hepatotoxicity in Liver Organoid Models
3. Discussion
4. Materials and Methods
4.1. DNA Constructs and Lentiviral Production
4.2. Cells and Cell Culture
4.3. Organoid Culture
4.4. Immunofluorescence (IF) Staining and Microscopy
4.5. Analysis of mRNA Expression
4.6. Western Blot Analysis
4.7. Analysis of Cell Proliferation and Apoptosis
4.8. Cell Motility and Contraction Assays
4.9. Hydroxyproline Assay
4.10. Prediction of MiRNA Targets and Dual-Luciferase® Reporter Assay
4.11. Lipid Droplet and Vitamin A Storage Assay
4.12. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Ariyachet, C.; Chuaypen, N.; Kaewsapsak, P.; Chantaravisoot, N.; Jindatip, D.; Potikanond, S.; Tangkijvanich, P. MicroRNA-223 Suppresses Human Hepatic Stellate Cell Activation Partly via Regulating the Actin Cytoskeleton and Alleviates Fibrosis in Organoid Models of Liver Injury. Int. J. Mol. Sci. 2022, 23, 9380. https://doi.org/10.3390/ijms23169380
Ariyachet C, Chuaypen N, Kaewsapsak P, Chantaravisoot N, Jindatip D, Potikanond S, Tangkijvanich P. MicroRNA-223 Suppresses Human Hepatic Stellate Cell Activation Partly via Regulating the Actin Cytoskeleton and Alleviates Fibrosis in Organoid Models of Liver Injury. International Journal of Molecular Sciences. 2022; 23(16):9380. https://doi.org/10.3390/ijms23169380
Chicago/Turabian StyleAriyachet, Chaiyaboot, Nattaya Chuaypen, Pornchai Kaewsapsak, Naphat Chantaravisoot, Depicha Jindatip, Saranyapin Potikanond, and Pisit Tangkijvanich. 2022. "MicroRNA-223 Suppresses Human Hepatic Stellate Cell Activation Partly via Regulating the Actin Cytoskeleton and Alleviates Fibrosis in Organoid Models of Liver Injury" International Journal of Molecular Sciences 23, no. 16: 9380. https://doi.org/10.3390/ijms23169380