Il 1β

Nitro blue tetrazolium chloride (N-BT) was purchased from the medicine supply station of the Academy of Military Medical Sciences (China). Lactate dehydrogenase (LDH), malondialdehyde (MDA), nitric oxide (NO), nitric oxide synthase (NOS), IL-1β, PI3K, and nuclear factor kB (NF-kB) kits were purchased from Nanjing Jiancheng Bioengineering Institute (China). High glucose Dulbecco's modified Eagle's medium (DMEM-H) and fetal bovine serum (FBS) were purchased from Gibco. MicroRotofor™ cell lysis kit and Bio-Plex Pro™ cell signaling MAPK panel were purchased from Bio-Rad (USA). Proteasome inhibitor, MG132, and IKK (inhibitor of nuclear factor kappa-B kinase, IκB) inhibitor, parthenolide, were purchased from Sigma.

Apelin 13 peptide was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Dulbecco’s modified Eagle’s medium (DMEM) was obtained from Hyclone (Logan, UT, USA). Fetal bovine serum (FBS) was obtained from Sijiqing Biotechnology (Hangzhou, China). Annexin V-FITC apoptosis detection kit was obtained from Kaiji Biotechnology (Hangzhou, China). 8-Oxo-dG, MDA, SOD, GSH, CAT, IL-1β, TNF-α, COX-2, and IL-6 detection kits were purchased from Jiancheng Biotechnology (Hangzhou, China). 2,3,5-triphenyltetrazolium chloride (TTC) and dihydroethidium (DHE) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Antibodies for P-AMPK, AMPK, GSK-3β, Nrf2, P-GSK-3β, P-ACC, ACC, NQO-1, HO-1, BCL-2, Bax, cleaved caspase 3, cleaved caspase 9, IP3, PLC, CaMKK, and β-actin were purchased from Cell Signaling Technology (Beverly, MA, USA). Annexin V-FITC apoptosis detection kit was purchased from Jiancheng Biotechnology (Hangzhou, China). All other chemicals used in this experiment were the purest grade commercially available.

XLVI (purity > 98.0%) and NPLC0393 (purity > 98.0%) were provided by Jiangsu Key Laboratory for Functional Substances of Chinese Medicine (Nanjing, China). Silymarin, CCK-8 kit, 0.25% trypsin, phenylmethylsulfonyl fluoride (PMSF), and NP-40 lysis buffer were obtained from Liangwei Biotechnology Co., Ltd. (Nanjing, China). The human LX2 cells were purchased from Mingzhoubio Company (No. MZ-0286). Fetal bovine serum (FBS), DMEM medium, penicillin-streptomycin (P/S) and phosphate-buffered saline (PBS) were obtained from Gibco Company (Grand Island, VT, USA). TGF-β1, antibodies recognizing COL1A1, α-SMA, p65, p-p65 (Ser536), GAPDH, and β-tubulin were obtained from Cell Signaling Technology (Danvers, MA, USA). Mouse secondary antibodies and rabbit secondary antibodies were provided by Signalway Antibody (College Park, MD, USA). The primers for COL1A1, α-SMA, and IL-1β were designed and synthesized by Shanghai Sangon Biological Engineering Co., Ltd. (Shanghai, China). ALT, AST, HYP, IL-1β, and TNF-α ELISA kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). RNase Mini Kits, Trizol reagent, and HiScript II Q RT SuperMix for qPCR were obtained from Vazyme Biotech Company (Nanjing, China).

2, 4, 6-trinitrobenzene sulfonic acid and sulfasalazine were bought from MP Biomedical (Aurora, OH, United States) and Yifeng Pharmacy (Nanjing, China), respectively. ELISA kits (IL-6, IL-1β, IFN-γ, and TNF-α) were purchased from Nanjing Jiancheng Bioengineering Institute Co., Ltd. CP were prepared according to our previous methods (Schicho et al., 2010 (link); Tao et al., 2017 (link)).

Before detection, cortical and hippocampal tissues were homogenized as before. TNF-α, IL-1β and IL-6 ELISA Kit Assays (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) were detected according to the manufacturer’s recommendations.