Glutathione peroxidase gsh px

The reference standard of n-alkanes (C₈−C₂₀) was received from ANPEL Laboratory Technologies (Shanghai) Inc. (Shanghai, China). 2,2-diphenyl-1-picrylhydrazyl (DPPH, 98%), 2,2′-amino-di(2-ethyl-benzothiazoline sulfonic acid-6) ammonium salt (ABTS, 98%), ascorbic acid (≥ 99%), 2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA), and acridine orange were acquired from Sigma-Aldrich (Shanghai, China). Malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) were obtained from Nanjing Jiancheng Biotechnology Co., Ltd (Nanjing, China). Dimethyl sulfoxide (DMSO) was obtained from MP Biomedicals (CA, USA). Diphenyl-1-pyrenylphosphine (DPPP, catalog no. DJ799) was obtained from Macklin (Shanghai, China). Other chemicals used in this study were of analytical grade.

Kaempferol (purity >98%) was obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China), and dissolved in dimethylsulfoxide (30 mg/ml), with a final concentration in phosphate buffer solution (PBS). SIRT1 inhibitor EX527 and peroxisome proliferator activated receptor gamma coactivator-1α (PGC-1α) inhibitor SR-18292 were bought from the Selleckchem (Houston, TX, United States), and both dissolved in dimethylsulfoxide (both were 30 mg/ml), with a final concentration in PBS. The antibodies of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X (Bax), cytochrome c (Cyt c) and cytochrome c oxidase subunit IV (COX IV) were all obtained commercially from Abcam (Cambridge, MA, United States). The detection kits of reactive oxygen species (ROS), malondialdehyde (MDA), total superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GSH-Px) were purchased from Jiancheng (Nanjing, JS, CHN). Annexin V-FITC apoptosis detection kit obtained from eBioscience (San Diego, CA, United States).

TTR extract (purity > 50%) was prepared by our laboratory. Sildenafil was purchased from Pfizer Pharmaceuticals Limited. SOD, glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) detection kits were obtained from Nanjing Jiancheng Bioengineering Institute. Enhanced RIPA lysate, broad-spectrum phosphatase inhibitor, and broad-spectrum protease inhibitor were obtained from Boster Bio Co., Ltd. BCA protein quantitation kits were purchased from Mei5 Biotech Co., Ltd. The following antibodies were used in the experiments: anti-Nrf2 (Wanleibio), anti-HO-1, anti-Bax, and anti-Bcl-2 (Abcam, USA), anti-β-actin (Beijing Affinity Biosciences), and HRP-conjugated AffiniPure goat anti-rabbit IgG (H + L) (Proteintech, Cat No. SA00001-2). Fetal calf serum was obtained from Gibco, USA. Primary endothelial cell basal culture medium, cell growth factor, and penicillin‒streptomycin double antibiotic were obtained from iCell Bioscience Inc. Pancreatin (0.25%) was purchased from HyClone, USA. Dimethyl sulfoxide was purchased from BIOFROXX, USA. Cell proliferation and toxicity detection kits were obtained from Bioss Biotech; apoptosis detection kits were obtained from USA BD, and reactive oxygen species (ROS) detection kits were obtained from Beijing Solarbio.

Lovastatin, lysophosphatidylcholine (LPC), hydrogen peroxide (H₂O₂), tumor necrosis factor alpha (TNFα), homocysteine thiolactone (HTL), D-glucose, mannitol, 1.1-dipheyl-2-picryl hydrazyl (DPPH), streptozotocin (STZ), acetylcholine, and phenylephrine were purchased from Sigma-Aldrich Company or Caymen chemical Company. Primary antibodies against PA200, p-AMPK, AMPK, and GAPDH, and secondary antibodies were obtained from Cell Signaling Company. Commercial kits malondialdehyde (MDA), superoxide dismutase (SOD) and Glutathione peroxidase (GSH-Px) were purchased from Jian-Cheng Bioengineering Institute (Nanjing, China). Oxidized low density lipoprotein (ox-LDL) was obtained from Guangzhou Yiyuan Biotech Co. Ltd. (Guangzhou, China). All drug concentrations are expressed as final working concentrations in the buffer.

Rat adrenal pheochromocytoma cells (PC12 cells; highly differentiated type) were purchased from the Cell Center of Shanghai Academy of Life Sciences, Chinese Academy of Sciences. Aβ_25-35 (purity ≥ 95%, no. Y0044) was from Beijing Biosynthesis Biotechnology Co., Ltd. (China). DMEM, streptomycin, trypsin, and fetal bovine serum were from HyClone Laboratories Inc. (Logan, Utah, USA). MTT was from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). A lactate dehydrogenase (LDH) kit was purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Superoxide dismutase (SOD), malondialdehyde (MDA), myeloperoxidase (MPO), and glutathione peroxidase (GSH-Px), as well as catalase (CAT), total antioxidant capacity (T-AOC), and acetylcholinesterase (AChE) kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The primary antibodies against sirtuin1 (Cat. No. 13161-1-AP), total Nrf2 (Cat. No. 16396-1-AP), nuclear Nrf2 (Cat. No. 66504-1-Ig), HO-1 (Cat. No. 66743-1-Ig), β-actin (Cat. No. 60008-1-Ig), and PCNA (Cat. No. 60097-1-Ig) were from Proteintech Co., Ltd. (Chicago, USA). HRP-labeled goat anti-mouse IgG antibody was purchased from Santa Cruz Biotechnology, Inc. (Cat. No. sc-2005; Santa Cruz, California, USA). HRP-labeled goat-rabbit IgG antibody was from Proteintech Co., Ltd. (Cat. No. 10285-1-AP; Chicago, USA).