Gsh px assay kit

After centrifugation (1,000 × g; 25°C; 5 min), the cell supernatant of each subgroup was collected to analyze the levels of MDA, GSH-Px, SOD and the inflammatory factors IL-6, IL-1β and TNF-α. The following kits were used according to the manufacturer's instructions: Micro MDA assay kit (cat. no. BC0020; Beijing Solarbio Science & Technology Co., Ltd.), GSH-Px assay kit (cat. no. BC1195; Beijing Solarbio Science & Technology Co., Ltd.), SOD activity detection kit (cat. no. BC0170; Beijing Solarbio Science & Technology Co., Ltd.), rat IL-1β ELISA kit (cat. no. RLB00; R&D Systems, Inc.), rat IL-6 Quantikine ELISA kit (cat. no. R6000B; R&D Systems, Inc.) and rat TNFα Quantikine ELISA kit (cat. no. RTA00; R&D Systems, Inc.).

Following treatment as described above, the cells were collected and centrifuged at 3,000 × g for 10 min at 4°C, and the supernatant was stored at −80°C. The LDH release levels, MDA levels and SOD and GSH-Px activity were detected using a LDH cytotoxicity kit (Beyotime Institute of Biotechnology), and MDA assay kit (Nanjing Jiancheng Institute of Bioengineering Institute), an SOD assay kit (Beijing Solarbio Science & Technology Co., Ltd.) and a GSH-Px assay kit (Beijing Solarbio Science & Technology Co., Ltd.), respectively. All assays were performed strictly according to the manufacturer's protocol.

The GSH-Px assay kit (Beijing Solarbio Science & Technology Co., Ltd.) was used to evaluate GSH-Px activity. HUVECs were collected and centrifuged at 8,000 × g for 10 min at 4°C after adding the extracting reagent of the GSH-Px assay kit. Subsequently, the corresponding reagents were added into the sample and fully mixed. The absorbance value was immediately determined at a wavelength of 412 nm according to the manufacturer's protocol.