Abstract:

Objective To investigate the effect of necrotic cells on oxidative stress of normal vascular smooth muscle cells, and explore the main active factors. Methods Necrosis of vascular smooth muscle cells was induced by glucose-free hypoxic conditions, and the supernatants were collected. Normal vascular smooth muscle cells were divided into control group (without any treatment), necrotic cell supernatants group (NCS group, treated with culture supernatants of necrotic cells) and NCS+interleukin 1 receptor antagonist (IL-1RA) group (treated with NCS+IL-1RA). The production of reactive oxygen species (ROS) of cells and the content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in culture supernatants were determined, and the expression of endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase(iNOS) and oxidase subunit p47phox mRNA and protein was detected by RT-PCR and Western blotting. Results The production of ROS and content of MDA in NCS group were significantly higher than those in control group (P<0.05), and the activity of SOD in NCS group was significantly lower than that in control group (P<0.05). The production of ROS and content of MDA in NCS+IL-1RA group was significantly lower than those in NCS group (P<0.05), and the activity of SOD in NCS+IL-1RA group was significantly higher than that in NCS group (P<0.05). RT-PCR and Western blotting revealed that the expression of iNOS, eNOS and p47phox mRNA and protein in cells in NCS group was significantly higher than that in control group (P<0.05), and the expression of iNOS, eNOS and p47phox mRNA and protein in cells in NCS+IL-1RA group was significantly lower than that in NCS group (P<0.05). Conclusion Necrotic cells can induce oxidative stress of normal vascular smooth muscle cells, and the inhibition of IL-1 signaling pathway can alleviate the oxidative stress.

Key words: vascular smooth muscle cells, necrosis, interleukin 1, oxidative stress