High glucose dmem

After obtaining informed consent from patients, we collected 30 pairs of HCC tissues and paraneoplastic tissues (from the Second Affiliated Hospital of Nanchang University), while one normal hepatocyte line (7702) and four HCC cell lines (97H, LM3, HepG2 and 7721) were cultured (cells were from the Shanghai Institute of Cell Biology). All cell lines were cultured in high glucose DMEM (Solarbio, Beijing, China) supplemented with 10% fetal bovine serum (bio-Industries, Beit-Haemek, Israel), 100 µg/ml streptomycin and 100 U/mL penicillin at 37 °C, and in a 5% CO2 humidified incubator.

H9C2 cells were cultured with 10% fetal bovine serum (Solarbio, Beijing, China), high-glucose-DMEM (Solarbio, Beijing, China) and antibiotics. GLPs were procured from SEITEbiogle (Chengdu, China). Angiotensin II was from Sigma (A9525, St. Louis, MO, United States). Antibodies against the following proteins were used: PPARγ (Abcam, ab272718, London, United Kingdom, 1:1000), PGC1α (Abcam, ab176328, London, United Kingdom, 1:1000), and β-actin (NCM Biotech, AB1020, Suzhou, Jiangsu, China, 1:1000). BCA protein assay kit was purchased from Beyotime (Shanghai, China), and T0070907 was obtained from Selleck (Houston, Texas, United States).

Penfluridol (purity ≥ 98.92%) was obtained from MedChemExpress (HY-B1077, Shanghai, China). Lipopolysaccharides (LPS from Escherichia coli 055: B5) was acquired from Sigma Chemical Co. (St. Louis, MO, USA). High-glucose DMEM and FBS were obtained from Solarbio (Beijing, China). Enhanced Cell counting Kit-8 (CCK-8) was purchased from Bioss (Beijing, China). Mouse and human ELISA kits for TNF-α, IL-6, IL-18, and IL-1β were acquired from BOSTER Biological Technology Co. Ltd., (Wuhan, China). The nitric oxide (NO), malondialdehyde (MDA), and superoxide dismutase (SOD) kits were obtained from Beyotime Institute of Biotechnology (Beijing, China). Primary antibodies against NLRP3, cleaved Caspase-1 and Nrf2 were purchased from Cell signaling. Antibodies against HO-1, lamin B, GAPDH, and β-tublin were purchased from proteintech Group, Inc (Wuhan, China). Primary antibody against IL-1β was purchased from Abcam (ab234437; Abcam, Cambridge, UK). HRP-Goat Anti-Mouse IgG (H + L) and HRP-Goat Anti-Rabbit IgG (H + L) were purchased from proteintech group, Inc (Wuhan, China).

Caco‐2 cells (BNCC350769, BeNa, Henan, China) were cultured in high glucose DMEM (Cat# 11 995; Solarbio, Inc.) containing 1% penicillin–streptomycin and 10% fetal bovine serum (FBS; Cat# 1611A; Bovogen, Australia) in a humid environment at 37 °C and 5% CO₂.