Regulatory Effect Of CCK-8 On LPS-induced Changes Of SOD Activity In VSMC And ECV-304

Objective: To investigate the regulatory effect of CCK-8 on LPS induced inducible SOD in VSMC and ECV-304.Methods: VSMC and Human vein endothelium line ECV-304 was cultured and incubated with LPS, CCK-8, CCK-8+ LPS, or solvent for different time. The cells were collected to assay SOD activity.Results:Part one (1) The effect of LPS on SOD activity and in VSMC: Incubation of VSMC with 0.5ug/ml LPS with the extension of incubating 2h, 6h, 16h. As compared with 6h group, the activity of SOD decrease in VSMC(P<0.01). (2) The effect of CCK-8 on SOD activity in VSMC: Incubation of VSMC with concentration 10-8mol/L of CCK-8 for 2h, 6h, 16h. As compared with 2h group, the activity of SOD increase in VSMC(P<0.05); As compared with 6h group, the activity of SOD increase in VSMC(P<0.01); As compared with 16h group, the activity of SOD increase in VSMC(P<0.05). (3) The effect of C＋L on SOD activity in VSMC: As compared with solvent 2h group, the activity of SOD decrease in VSMC(P<0.01); As compared with LPS0.5ug/ml 2h group, the activity of SOD decrease in VSMC(P<0.01); As compared with CCK-810-8mol/L 2h group, the activity of SOD decrease in VSMC(P<0.01); As compared with LPS0.5ug/ml 6h group, the activity of SOD decrease in VSMC(P<0.01); As compared with LPS0.5ug/ml 16h group, the activity of SOD decrease in VSMC(P<0.01); As compared with CCK-810-8 mol/L 16h group, the activity of SOD decrease in VSMC(P<0.01).Part two (1) The effect of LPS on SOD activity and in ECV-304: Incubation of ECV-304 exposed to LPS with different concentration (0.01ug/ml, 0.1ug/ml and 1ug/ml)with the extension of incubating 2h, 6h, 16h. As compared with solvent 2h group, the activity of SOD decrease in ECV-304, LPS 0.01ug/ml(P<0.01), LPS 0.1ug/ml(P<0.01)and LPS 1ug/ml(P<0.01); As compared with solvent 6h group, the activity of SOD decrease in ECV-304,LPS 0.01ug/ml(P<0.01), LPS 0.1ug/ml(P<0.01); As compared with solvent 16h group, the activity of SOD decrease in ECV-304,LPS 0.01ug/ml(P<0.01), LPS 1ug/ml(P<0.05). (2) The effect of CCK-8 on SOD activity in ECV-304: Incubation of ECV-304 with different concentration 10-9mol/L, 10-8mol/L of CCK-8 for 2h, 6h, 16h. As compared with solvent 2h group, the activity of SOD decrease in CCK-810-8mol/L(P<0.01); As compared with solvent 6h group, the activity of SOD increase in CCK-810-9mol/L(P<0.01); The activity of SOD decrease in CCK-810-8mol/L(P<0.01)；As compared with solvent 16h group, the activity of SOD increase in CCK-810-8mol/L(P<0.01). (3) The effect of C＋L on SOD activity in ECV-304: Incubation of ECV-304 with CCK-810-8mol/L＋LPS 0.1ug/ml with the extension of incubating 2h, 6h, 16h. As compared with solvent 2h group, the activity of SOD increase in ECV-304(P<0.05); As compared with LPS0.1ug/ml 2h group, the activity of SOD increase in ECV-304(P<0.01)；As compared with CCK-8 10-8mol/L 2h group,the activity of SOD increase in ECV-304(P<0.01)；As compared with solvent 6h group, the activity of SOD increase in ECV-304(P<0.01)；As compared with LPS 0.1ug/ml 6h group,the activity of SOD increase in ECV-304(P<0.01); As compared with solvent 16h group, the activity of SOD increase in ECV-304(P<0.01); As compared with LPS 0.1ug/ml 16h group,the activity of SOD increase in ECV-304(P<0.01); As compared with CCK-8 10-8mol/L 16h group,the activity of SOD decrease in ECV-304(P<0.01). (4) The effect of LPS on Cu/Zn-SOD activity and in ECV-304: Incubation of ECV-304 exposed to LPS with different concentration (0.01ug/ml, 0.1ug/ml and 1ug/ml) with the extension of incubating 2h, 6h, 16h. As compared with solvent 6h group, the activity of Cu/Zn-SOD increase in ECV-304, LPS 0.1ug/ml(P<0.05), LPS 1ug/ml(P<0.01); As compared with solvent 16h group, the activity of Cu/Zn-SOD increase in ECV-304,LPS0.01ug/ml(P<0.01), LPS 0.1ug/ml(P<0.01)and LPS 1ug/ml(P<0.01). (5) The effect of CCK-8 on Cu/Zn-SOD activity in ECV-304: Incubation of ECV-304 with with different concentration 10-9mol/L, 10-8mol/L of CCK-8 for 2h, 6h, 16h. As...