Caspase 3

Curcumin (CUR) and THC were obtained from Wuhan reagent company (Wuhan, China). Dimethylsulfoxide (DMSO), 5-fluoro-2,4 (1 h,3 h) pyrimidinedione (5-FU), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), rhodamine 123 (Rh123), ethylenediaminetetraacetic acid (EDTA), RNAse-A and propidium iodide (PI), phenylmethyl-sulfonyl fluoride (PMSF), 4′,6-diamidino-2-phenylindole (DAPI), electro-chemi-luminescence (ECL), dichlorofluorescein diacetate (DCFH-DA), and tris-HCl and glycine were obtained from Sigma-Aldrich (St, Louis, MO, USA). Assay kits of BCA, caspase-3, caspase-9, and lactate dehydrogenase (LDH) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Phosphate buffer solution (PBS), 0.1% tween-20 in phosphate buffer solution (PBST), and sodium dodecyl sulfonate (SDS) were purchased from Zhengjiang Wanbang Pharmaceutical Co. (Wenling, China). The primary antibodies against Bax, Bcl-2, poly (ADP-ribose) polymerase (PARP), cytochrome c, and the horseradish peroxidase (HPR)-conjugated goat anti-mouse secondary antibody were provided by BioVision, Inc., (BioVisio, Palo Alto, CA, USA). All the other cell culture reagents were purchased from Sinopharm (Beijing, China), and all the other chemicals were of analytical grade.

All laboratory animal procedures were conducted in strict accordance with the National Institutes of Health Guidelines for the Use of Laboratory Animals. All procedures involving animals and their care have been approved by the Animal Ethics Committee of the Yijishan Hospital of Wannan Medical College (SCXK 2019-0007). All the mice were male between 8 and 12 weeks of age. The male mice were placed under standard conditions, with a 12-hour light/dark cycle and plenty of water and food. All efforts were made to minimize suffering. Male Balb/c mice were purchased from the Shanghai Laboratory Animal Center (Shanghai, China). Lonicera japonica was purchased from Zhongshan chemist's shop in Wuhu, China. Standard monosaccharides, including D-mannose, D-rhamnose, D-glucose, D-fucose, D-xylose, D-galactose, and D-arabinose were bought from Sigma (St. Louis, MO, USA). Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), dimethyl sulfoxide (DMSO), and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Annexin V-FITC was purchased from BioLegend, Inc. (San Diego, CA). AST, CPK, LDH, Caspase-3, Caspase-8, and Caspase-9 kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). SOD, CAT, GSH-Px, and MDA kits were purchased from BOMEI Biotechnology Co., Ltd. (Hefei, China).

Butin was purchased from National Institute for the Control of Pharmaceutical and Biological Products, and the purity being more than 98%. Dulbecco’s Modified Eagle’s Medium (DMEM) and other cell culture supplies were purchased from Gibco (Life Technologies, Grand Island, NY). 3-[4, 5-Dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) and 2, 3, 5-triphenyltetrazolium chloride (TTC) were obtained from Sigma (St. Louis, MO). The detection kits of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH) and glutathione disulfide (GSSG) and caspase 3 were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, P.R. China). Monoclonal rabbit anti-mouse P-AMPK, AMPK, P-GSK-3β, GSK-3β, Bcl-2, β-actin, Bax, NQO1, HO-1 and Nrf2 antibodies were purchased from Cell Signaling Technologies, MA, USA. All other chemicals used in this experiment were the purest grade commercially available.