Total RNA was extracted from the livers and hepatocytes using TRIzol™ Reagent (#15596018; ThermoFisher Scientific) and reversely transcribed to cDNA by the RT First Strand cDNA Synthesis Kit (#G3330; Servicebio, Wuhan, China) according to the manufacturer's instructions. Next, quantitative real-time PCR was performed with SYBR Green Master Mix and normalized to the internal control using the 2-∆∆Ct method [36 (link)–39 (link)]. The thermocycling conditions were as follows: 95°C for 10, then 40 cycles of 95°C for 2 sec, 60°C for 20 sec, and 70°C for 10 sec. The primer sequences were listed as follows: collagen 1α1 (Col1α1), forward, 5′-AGGCTTCAGTGGTTTGGATG-3′ and reverse, 5′-CACCAACAGCACCATCGTTA-3′; Col3α1, forward, 5′-CCCAACCCAGAGATCCCATT-3′ and reverse, 5′-GAAGCACAGGAGCAGGTGTAGA-3′; connective tissue growth factor (CTGF), forward, 5′-TGTGTGATGAGCCCAAGGAC-3′ and reverse, 5′-AGTTGGCTCGCATCATAGTTG-3′; transforming growth factor-beta 1 (TGF-β1), forward, 5′-TGCGCTTGCAGAGATTAAAA-3′ and reverse, 5′-CGTCAAAAGACAGCCACTCA-3′; miR-137-3p, forward 5′-TTATTGCTTAAGAATACGCG-3′ and reverse, 5′-TCGTATCCAGTGCAGGGTC-3′; GAPDH, forward, 5′-CGTGCCGCCTGGAGAAACC-3′ and reverse, 5′-TGGAAGAGTGGGAGTTGCTGTTG-3′; and U6, forward, 5′-CTCGCTTCGGCAGCACA-3′ and reverse, 5′-AACGCTTCACGAATTTGCGT-3′.
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