The Exploration Of The Mechanism MgSO₄ Prevent And Treat The Radiation-Induced HUVEC Injury

Objective:To investigate the effect and possible mechanism of MgSO4 on cells’ survival rate,DNA damage and inflammatory response in HUVEC induced by X-ray.Methods:1. CCK-8 assay was adopted to examine the influence of MgSO4 on the cells’ survival rate;Clonogenic assay was applied to observe the cells’ proliferation in order to evaluate the radiation toxicity reduced by MgSO4.2. laser confocal microscope was used to test the γ-H2 AX foci formation;the flow cytometry and western blot was employed to measure the γ-H2 AX expression.3. Real-time PCR assay was used to detect the ICAM-1 、 NF-κB mRNA expression;laser confocal microscope and flow cytometry was taken to observe the ICAM-1 expression; laser confocal microscope and western blot was applied to examine the NF-κB expression.Results:1. CCK-8 assay results showed that MgSO4 with the concentration of 1.25mg/mL to 6.25 mg/mL was no-toxic to HUVEC,and the MgSO4 with concentration of1.25 mg/mL could improve the survial rate of the HUVEC treated with X-ray,and it showed that MgSO4 would inhibit the cell’s survival rate,when concentration increased to 12.5 mg/mL; Clonogenic assay results suggested that 1.25 mg/mL to 2.5 mg/mL concentration of MgSO4 could increase the cell’s colony forming efficiency while 12.5mg/mL could inhibit it.2. Immunofluorescent analysis、Flow cytometry and Western blot demonstratedthat γ-H2 AX foci achieve the highest level in 30 min to 1 h after radiation and foci number was decreased in time-dependent manners,MgSO4 could significantly reduce the foci formation and γ-H2 AX protein increased.3. Real-time PCR test results showed that ICAM-1、NF-κB mRNA expression was increased after radiation and they both inhibited by 1.25 mg/mL MgSO4; aser confocal microscope and flow cytometry demonstrated that MgSO4 could inhibite ICAM-1 protein express;Given to 4 Gy X-ray,NF-κB in HUVEC translocate from cytoplasm to nuclear,MgSO4 significantly inhibited the translocation and activation of NF-κB.Conclusions:1. A certain concentration of the MgSO4 could increase cells’ survival rate and reduced the DNA damage after radiation.2. MgSO4 down-regulation the expression of ICAM-1 via inhibited the translocation and activation of NF-κB.