Nf κb

Lipopolysaccharide (LPS, Escherichia coli 0111:B4) was purchased from Sigma (St. Louis, USA). ELISA rat IL-6 and TNF-α kits were purchased from Hefei Bomei Biotechnology CO., LTD (Hefei China). Primary polyclonal antibodies β-actin, HO-1, COX-2, NF-κB, p-NF-Κb, p-ERK1/2, ERK were purchased from Bio Basic Inc., Canada.

Hearts stored at − 70 °C were lysed and homogenized inice-cold lysis buffer (50 mmol/L HEPES, 100 mmol/L sodium pyrophosphate, 10 mmol/L sodium orthovanadate, 100 mmol/L sodium fluoride, 10 mmol/L EDTA, and 1% Triton X-100) containing 2 mmol/L phenylmethyl sulfonyl fluoride. Homogenates were centrifuged at 12,000 g for 20 min at 4 °C. Equal amounts of denatured protein in supernatants were separated by electrophoresis on 12% SDS-PAGE and then transferred onto nitrocellulose membranes. The membranes were incubated with rabbit anti-rat primary antibodies such as β-actin, HO-1, COX-2, NF-κB, p-NF-κB, p-ERK1/2, ERK, (Bio Basic Inc., Canada) overnight at 4 °C. Subsequently, membranes were incubated with a horseradish peroxidase-conjugated secondary goat anti-rabbit antibody. After rinsing, proteins were visualized by DAB staining (Bio Basic Inc., Canada).

Lycopene (502-65-8, 98% purity) was purchased from Nanjing Xinkailong Bioengineering Co., Ltd (Nanjing, China). Carbon tetrachloride (CCl₄) was purchased from Suzhou Baiyu Chemical Co., Ltd (Suzhou, China). Antibodies β-actin, heme oxygenase 1 (HO-1), IκB α, p-IκB α, NF-κB, and p-NF-κB were obtained from Bio Basic Inc. (Markham, ON, Canada). Antibodies SIRT 1, peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α), microtubule-associated protein 1 light chain 3B (LC3), P62, Beclin 1, regulated in development and DNA damage responses 1 (REDD1), Src homology 2 domain-containing phosphatase 2 (SHP2), transforming growth factor beta (TGF-β), and α-SMA were purchased from Abcam (Cambridge, MA, USA).