Effect of Ad. mTGF-1-gene transfection in vitro combined with FTY720 administration on ischemia-reperfusion injury in rat cardiac allografts

Abstract

Objective To explore the role of Ad. mTGF-β1 gene transfection in vitro combined with FTY720 administration on ischemia-reperfusion injury after heart transplantation in rats, to determine if the combination of them had a synergistic effect, and to discuss the mechanisms. Methods All animals were divided into 5 groups:control group,control vector only-group, FTY720-treated group, Ad. mTGF-β1-transfected group,and Ad. mTGF-β1 FTY720-treated group. All recipients were sacrificed, and the donor hearts were removed at the 8th h after transplantation. The expression of mTGF-β1,ICAM-1,and NF-κB in the heart allografts was detected by using immunohistochemieal staining. Gene product expression in the tissues was quantified by one step RT-PCR. SOD,MDA,and MPO in the grafts were measured. Results The expression of mTGF-β1 in Ad. mTGF-β1-transfected and Ad. mTGF-β1 FTY720-treated groups was (1.08±0.24) and (1.16±0.22), higher than in other three groups (P<0.01). The expression of ICAM and NF-κB in Ad. mTGF-β1-transfected group was (2.43±0.46) and (9.80±1.85), and (1.90±0.20) and (10.1±2.27) in Ad. mTGF-β1 FTY720-treated group respectively,which were lower than in other three groups (P<0.01). The apoptotic index in FTY720 treated, Ad. mTGF-β1-transfected,and Ad. mTGF-β1 FTY720-treated groups was (9.2±1.12), (5.9±1.09) and (5.4±0.77) respec-tively, which was significantly lower than in other groups (P<0.01). SOD activity in FTY720, Ad. mTGF-β1-transfected, and Ad. mTGF-β1 FTY7 20-treated groups was (51.0 3±5.54), (55.91±6.66), and (73.42±6.42) U/mg respectively, higher than other groups (P<0.01). MDA content in FTY720, Ad.mTGF-β1-transfected, and Ad. mTGF-β1 FTY720-treated groups was (10.90±1.93), (11.02±2.45),and (9.28±1.64) U/g respectively, lower than in other groups (P<0.01). MPO activity in FTY720,Ad. mTGF-β1-transfected, and Ad. mTGF-β1 FTY720-treated groups was (4.38±1.43), (4.63±1.04),and (3.16±0.64) U/g respectively, lower than in other groups (P<0.01). Interactions were proven above in gene transfer and FTY720. Conclusion Combined Ad. mTGF-β1 gene transfection in vitro with FTY720 ameliorates the myocardial ischemia-reperfusion injury in rat heart transplantation, in-creases the SOD activity and decreases the MDA content and MPO activity, and reduce the expressions of ICAM-1 and NF-κB. As a result,the combination of mTGF-β1 gene transfection in vitro and FTY720 ad-ministration showed an additive effect. Key words: Heart transplantation; TGF-β1; FTY720; Reperfusion Injury