Introduction
Materials and methods
Reagents
Cell culture and grouping
Cell transient transfection and exposure to hyperoxia
Real-time polymerase chain reaction (RT-PCR)
Name | Primer sequence | Product |
---|---|---|
C/EBPα | Forward: 5′-GGTGGACAAGAACAGCAACG-3′ | 124 bp |
Reverse: 5′-GGTCAGCTCCAGCACCTTCT-3′ | ||
SP-C | Forward: 5′- TTACCACTGCCACCTTCTCC-3’ | 139 bp |
Reverse: 5′-TCAAGACTGGGGATGCTCTC-3’ | ||
β-actin | Forward: 5′-GCAGAAGGAGATTACTGCCCT-3’ | 136 bp |
Reverse: 5′-GCTGATCCACATCTGCTGGAA-3’ |
Western blot
Cell proliferation
Cell cycle analysis
Cell apoptosis analysis
Statistical analysis
Results
pcDNA3.1(+)-C/EBPα induces increased C/EBPα protein expression
Overexpression of C/EBPα improves the secretion of SP-C in AEC II after exposure to hyperoxia
Overexpression of C/EBPα promotes cell proliferation after exposure to hyperoxia
Group | OD value |
---|---|
Air (AG) | 1.91 ± 0.63 |
Air+pcDNA3.1(+)-C/EBPα (A + C) | 1.89 ± 0.52& |
Air+ pcDNA3.1(+)-empty (A + E) | 1.87 ± 0.46 |
Hyperoxia (HG) | 0.89 ± 0.44* |
Hyperoxia+pcDNA3.1(+)- C/EBPα (H + C) | 2.43 ± 0.25# |
Hyperoxia+pcDNA3.1(+)-empty (H + E) | 0.76 ± 0.32* |
Overexpression of C/EBPα regulates cell cycle distribution after hyperoxia exposure
Cell | G1 (%) | S (%) | G2 (%) |
---|---|---|---|
Air (AG) | 68.61 ± 2.68 | 28.19 ± 2.45 | 4.17 ± 1.87 |
Air+pcDNA3.1(+)- C/EBPα (A + C) | 64.92 ± 3.12& | 28.11 ± 2.53& | 7.51 ± 2.98& |
Air+pcDNA3.1(+)-empty (A + E) | 67.26 ± 2.56 | 29.77 ± 1.98 | 6.02 ± 2.73 |
Hyperoxia (HG) | 91.09 ± 2.41* | 5.95 ± 0.98* | 3.00 ± 1.61* |
hyperoxia+pcDNA3.1(+)-C/EBPα (H + C) | 51.15 ± 3.36# | 37.61 ± 2.76# | 10.29 ± 2.02# |
Hyperoxia+pcDNA3.1(+)-empty (H + E) | 92.29 ± 1.97* | 6.41 ± 3.17* | 2.01 ± 1.91* |
Overexpression of C/EBPα decreases cells apoptosis and necrosis after exposure to hyperoxia
Group | Apoptosis rate |
---|---|
Air (AG) | 9.76 ± 0.67 |
Air+pcDNA3.1(+)-C/EBPα (A + C) | 8.97 ± 0.46& |
Air+pcDNA3.1(+)-empty (A + E) | 9.36 ± 0.39 |
Hyperoxia (HG) | 13.01 ± 0.86* |
Hyperoxia+pcDNA3.1(+)-C/EBPα (H + C) | 9.51 ± 0.62# |
Hyperoxia+pcDNA3.1(+)-empty (H + E) | 13.88 ± 0.54* |