Caspase 8

The activities of 8-hydroxy-2′-deoxyguanosine (8-OHdG), caspase-3, caspase-8, caspase-9 (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), aspartate aminotransferase transaminase (AST) and alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma glutamyltransferase (GGT) (Shanghai Yuanye Bioengineering Institute, Shanghai, China) were determined using commercially available enzyme-linked immune sorbent assay (ELISA) kits. Briefly, liver tissues were homogenized in 0.9% of saline solution and then centrifuged at 12,000 ×g for 15 min, to release the enzymes into the solution. Then, the microplates were coated of with anti-8-OHdG, caspase-3, caspase-8, caspase-9, AST, ALT, GGT, and ALP, followed by detection with a horseradish peroxidase-labeled substrate after incubation for 10 minutes at 37°C. Absorbance values were then read in a spectrophotometer at 450 nm.

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).