Total RNA of liver tissues was isolated with the RNeasy Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. Eight hundred nanograms total RNA was reverse-transcribed into cDNA using the PrimeScript RT reagent Kit (Takara, Tokyo, Japan) following the manufacturer’s instructions. The PCR amplification products were quantified by SYBR-green based qRT-PCR (Takara) following a standard procedure. The mRNA expression levels of target genes were normalized by β-Actin. Primer sequences are provided in Table 1 .
Primers Used for qRT-PCR
| Gene | Sequence 5′—3′ forward | Sequence 5′—3′ reverse |
|---|---|---|
| β-Actin | GTGACGTTGACATCCGTAAAGA | GCCGGACTCATCGTACTCC |
| Il-6 | GCTACCAAACTGGATATAATCAGGA | CCAGGTAGCTATGGTACTCCAGAA |
| Il-1β | TGTAATGAAAGACGGCACACC | TCTTCTTTGGGTATTGCTTGG |
| Tnfα | TTCTATGGCCCAGACCCTCA | TTTGCTACGACGTGGGCTAC |
| Dj-1 | GTGCAGTGTAGCCGTGATGT | CCTCCTGGAAGAACCACCAC |