Construction of hypoxia/reoxygenation injury model in Caco-2 cell monolayer
Objective To construct the injury model of the monolayer of Caco-2 cells,and to explore the effect of carbon monoxide releasing molecule 3(CORM-3)on the injury model of Caco-2 cells.Methods Caco-2 cells were cultured and using 5%CO2,1%O2 and 94%N2 at 37 degrees incubator to develop a hypoxia/reoxygenation(H/R)model.According to the hypoxia time,Caco-2 cells were divided four groups,blank control group,H/R 1 group(hypoxia for 4 hours and reoxygenation for 4 hours),H/R two group(hypoxia for 8 hours and reoxygenation for 4 hours),and H/R three group(hypoxia for 12 hours and reoxygenation for 4 hours).Dissolve CORM-3 drug powder and prepare inactive CORM-3(iCORM-3),which was divided into 6 groups according to drug concentration,blank control group(Control),hypoxic reoxygenation injury model(H/R),H/R+300 μmol/L CORM-3(H/R+C1),H/R+400μmol/L CORM-3(H/R+C2),H/R+500μmol/L CORM-3(H/R+C3),and H/R+500μmol/L iCORM-3(H/R+C4).The cell counting kit 8(CCK8)kit was mainly used to detect the cell viability,and the morphological changes were ob-served under an inverted microscope.Fluorescein sodium was used to measure the permeability of the Caco-2 cell monolayer.Results With the prolongation of hypoxia time,changes in the morphology of Caco-2 cells were observed,with the disappearance of inter-cellular connections,a decrease in cell viability,and an increase in cell monolayer permeability.After receiving CORM-3 drug intervention treatment,compared to the untreated injury group,Caco-2 cells showed varying degrees of repair,and cell viability increasing with the in-crease of drug concentration.Conclusion This study successfully constructs a damage model of Caco-2 cell monolayer hypoxia reoxygen-ation,and found that CORM-3 may have a protective effect on the damage model of Caco-2 cell monolayer hypoxia reoxygenation.