Introduction
Methods
Ethical considerations
Cell culture
Immunocytochemistry
Propofol and coenzyme Q10 treatment
Cellular ATP levels assay
Cell viability and caspase 3/7 activity assay
Measurement of lactate concentrations in culture media
NAD+/NADH ratio assay
Mitochondrial membrane potential assay
Mitochondrial morphologic assessment
RNA extraction
Analyses for mRNA expression by reverse transcription PCR
Gene | Primer | Sequence (5′–3′) |
---|---|---|
ACTB
| Left | ccaaccgcgagaagatga |
Right | ccagaggcgtacagggatag | |
NDUFS8
| Left | tgcttggcaaggcaagtag |
Right | tctcgggatcctgcatgt | |
SDHB
| Left | ggggcctgcagttcttatg |
Right | aggcgctcctctgtgaagt | |
PGC-1alpha | Left | tgagagggccaagcaaag |
Right | ataaatcacacggcgctctt | |
NRF1
| Left | ccatctggtggcctgaag |
Right | gtagtgcctgggtccatga | |
NRF2
| Left | ggacgggtctaggtgagaca |
Right | tggctggagtatttcaaaggat | |
PDSS1
| Left | agatctggggtgaaaagaagg |
Right | ccaattcgtgccagagcta | |
PPARalpha
| Left | gcactggaactggatgacag |
Right | tttagaaggccaggacgatct | |
CPT1B
| Left | tgtgagtgactggtgggaag |
Right | ttgatgagcacaaggtccat |
Statistical analysis
Results
Induced pluripotent stem cell-derived cardiomyocyte phenotypes and responses to toxic effects of high concentrations of propofol
Concentration-dependent induction of mitochondrial dysfunction by propofol
Gene | Expression ratio (vs. control) | 95% CI |
---|---|---|
Complex I | ||
NDUFB2 | 0.83 | 0.69, 0.96 |
NDUFB3 | 0.82 | 0.69, 0.94 |
NDUFB4 | 0.80 | 0.67, 0.92 |
NDUFB9 | 0.84 | 0.72, 0.95 |
NDUFB10 | 0.85 | 0.72, 0.98 |
NDUFS3 | 0.80 | 0.63, 0.97 |
NDUFS8 | 0.59 | 0.47, 0.71 |
NDUFV2 | 0.84 | 0.75, 0.94 |
NDUFV3 | 0.63 | 0.46, 0.80 |
Complex II | ||
SDHB | 0.83 | 0.74, 0.92 |
SDHD | 0.48 | 0.22, 0.74 |
Complex III | ||
UQCRC1 | 0.81 | 0.64, 0.98 |
UQCRC2 | 0.84 | 0.69, 0.99 |
UQCRFS1 | 0.86 | 0.75, 0.97 |
Complex IV | ||
COX4I1 | 0.65 | 0.45, 0.85 |
COX4I2 | 0.59 | 0.26, 0.91 |
COX8C | 0.82 | 0.67, 0.96 |
Complex V | ||
ATP5G1 | 0.81 | 0.66, 0.96 |
ATP6V0D2 | 0.30 | 0.11, 0.48 |
OXA1L | 0.76 | 0.64, 0.88 |