Bleomycin-Induced Pulmonary Fibrosis in Transgenic Mice Carrying the Human MUC5B rs35705950 Variant
Abstract
:1. Introduction
2. Materials and Methods
2.1. Experimental Animals
2.2. Generation of Human MUC5B [rs35705950] Transgenic Mice
2.3. Pulmonary Fibrosis Induction
2.4. Collection of BALF and Peripheral Blood
2.5. Lung Tissue Collection
2.6. Biochemical Analysis
2.7. Immunohistochemical Staining
2.8. Gene Expression Analysis
2.9. Statistical Analysis
3. Results
3.1. Development of a Transgenic Mouse Model Expressing the Human MUC5B rs35705950 with the Mutant T Allele
3.2. Significant Expression of MUC5B in the Proximal Bronchial Segments in Human MUC5B rs35705950 Transgenic Mice with Lung Fibrosis
3.3. Reduced Infiltration of Inflammatory Cells in Human MUC5B rs35705950 Transgenic Mice with Lung Fibrosis
3.4. Reduced Expression of Inflammatory Cytokines in Human MUC5B rs35705950 Transgenic Mice with Lung Fibrosis
3.5. Decreased Expression of Growth Factors in Human MUC5B rs35705950 Transgenic Mice with Lung Fibrosis
3.6. Reduced Expression of Extracellular Matrix Markers in Human MUC5B rs35705950 Transgenic Mice with Lung Fibrosis
3.7. Decreased Lung Fibrosis in Human MUC5B rs35705950 Transgenic Mice
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gene & Direction | Sequence (5′ to 3′) | Length (nt) | Tm (°C) | Accession Number | Position | Product |
---|---|---|---|---|---|---|
hMUC5B | ||||||
Forward | GCCCACATCTCCACCTATGAT | 21 | 61.1 | NM_002458.3 | 1349-1369 | 141 bp |
Reverse | GCAGTTCTCGTTGTCCGTCA | 20 | 62.4 | 1489-1470 | ||
mMuc5B | ||||||
Forward | TCCCTAGCATGAGCGCCTTA | 20 | 62.6 | NM_028801.2 | 197-216 | 178 bp |
Reverse | CCACGACGCAGTTGGATGTT | 20 | 63 | 374-355 | ||
mPeriostin | ||||||
Forward | CACGGCATGGTTATTCCTTCA | 21 | 60.4 | NM_001198766.1 | 547-567 | 151 bp |
Reverse | TCAGGACACGGTCAATGACAT | 21 | 61.1 | 697-677 | ||
mFibronectin | ||||||
Forward | TTCAAGTGTGATCCCCATGAAG | 22 | 60 | NM_010233.2 | 7126-7147 | 154 bp |
Reverse | CAGGTCTACGGCAGTTGTCA | 20 | 61.5 | 7279-7260 | ||
mCollagen I | ||||||
Forward | TAAGGGTCCCCAATGGTGAGA | 21 | 63.8 | NM007742.4 | 107-127 | 203 bp |
Reverse | GGGTCCCTCGACTCCTACAT | 20 | 60.3 | 309-290 | ||
mα-SMA | ||||||
Forward | CAGGATGCAGAAGGAGATCAC | 21 | 60.7 | NM007392.2 | 1009-1029 | 364 bp |
Reverse | TGTTGCTAGGCCAGGGCTAC | 20 | 62.6 | 1372-1353 | ||
mGapdh | ||||||
Forward | TGGCCTTCCGTGTTCCTAC | 19 | 61.3 | NM_008084.4 | 686-704 | 178 bp |
Reverse | GAGTTGCTGTTGAAGTCGCA | 20 | 60.9 | 863-844 | ||
Gapdh: glyceraldehyde 3-phosphate dehydrogenase; αSMA, αsmooth muscle actin; h, human; m, mouse. |
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Tharavecharak, S.; Fujimoto, H.; Yasuma, T.; D’Alessandro-Gabazza, C.N.; Toda, M.; Tomaru, A.; Saiki, H.; Uemura, M.; Kogue, Y.; Ito, T.; et al. Bleomycin-Induced Pulmonary Fibrosis in Transgenic Mice Carrying the Human MUC5B rs35705950 Variant. Cells 2024, 13, 1523. https://doi.org/10.3390/cells13181523
Tharavecharak S, Fujimoto H, Yasuma T, D’Alessandro-Gabazza CN, Toda M, Tomaru A, Saiki H, Uemura M, Kogue Y, Ito T, et al. Bleomycin-Induced Pulmonary Fibrosis in Transgenic Mice Carrying the Human MUC5B rs35705950 Variant. Cells. 2024; 13(18):1523. https://doi.org/10.3390/cells13181523
Chicago/Turabian StyleTharavecharak, Suphachai, Hajime Fujimoto, Taro Yasuma, Corina N. D’Alessandro-Gabazza, Masaaki Toda, Atsushi Tomaru, Haruko Saiki, Mei Uemura, Yurie Kogue, Toshiyuki Ito, and et al. 2024. "Bleomycin-Induced Pulmonary Fibrosis in Transgenic Mice Carrying the Human MUC5B rs35705950 Variant" Cells 13, no. 18: 1523. https://doi.org/10.3390/cells13181523