Acetylcysteine nac

The LX-2 human HSC line (Sigma, St.Louis, MO, USA, SCC064) was cultured in Dulbecco’s modified Eagle’s medium-high glucose (DMEM; Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS, Sigma, St.Louis, MO, USA, F8318-500Ml) and 1% penicillin/streptomycin (Gibco, Waltham, MA, USA , 15140-122). The cells were incubated at 37 °C with 5% CO₂. For the experiment, LX-2 cells were treated with various concentrations (10, 50 µM) of 3-Indolepropionic acid (IPA, Sigma, St.Louis, MO, USA, 220027-1G) for 24 h. Dimethylsulfoxide (DMSO, MP Biomedicals, Irvine, CA, USA, 67-68-5) was used as the control. Additionally, TGFβ1 treatment with a concentration of 5 ng/mL (Peprotech, Rocky Hill, NJ, USA, 100-21) was used as a positive control. In terms of inhibitors, the ROS inhibitor Acetylcysteine (NAC, Selleck, Houston, TX, USA, S1623) was pre-incubated at a concentration of 5 mM for 2 h before adding IPA. The p38 pathway inhibitor SB 202190 (p38i, MCE, Monmouth Junction, NJ, USA, HY-10295) was pre-incubated at a concentration of 20 µM for 2 h before adding IPA. The JNK pathway inhibitor SP600125 (JNKi, MCE, Monmouth Junction, NJ, USA, HY-12041) was pre-incubated at a concentration of 10 µM for 2 h before adding IPA.