干扰素-γ对兔动脉损伤后平滑肌细胞增殖和迁移的作用

Role of interferon-γ on smooth muscle cells proliferation and migration after balloon injury

目的 探讨干扰素 -γ(IFN-γ)对动脉损伤后平滑肌细胞 (SMC)增殖和迁移的作用及其作用机制。方法 建立兔髂动脉球囊损伤动物模型 ,培养损伤后动脉 SMC,实验分 4组 :对照组、转化生长因子 -β1(TGF-β1)组、IFN-γ组、TGF-β1+ IFN-γ组。每组细胞加培养液或加入 10μg/L TGF-β1或 (和 ) 50 0 k U/L IFN-γ刺激 72 h,细胞计数和四甲基偶氮唑盐 (MTT)法测定各组 SMC的增殖 ,同时测定 SMC迁移。明胶酶谱分析 SMC中基质金属蛋白酶 -2 (matrix metalloproteinase-2 ,MMP-2 ,66k D蛋白 )的活力。结果 与对照组比较 ,72 h后 TGF-β1组细胞数和迁移距离增加 (P <0 .0 1) ,MTT A值检测细胞增殖抑制率为 -19.4% ;IFN-γ组细胞数和迁移距离减少(P <0 .0 1) ,细胞增殖抑制率为 15.8% ;TGF-β1+ IFN-γ组细胞数和迁移距离低于 TGF-β1组 ,细胞增殖抑制率为 -9.1%。明胶酶谱分析显示各组细胞均可检测到 MMP-2。,TGF-β1组还可检测 MMP-2酶原 (pro-MMP-2 ,72k D蛋白 )的表达。与对照组 (10 0 % )比较 ,TGF-β1组 MMP-2相对活力为 14 3 % ,IFN-γ组为 95% ,TGF-β1+ IFN-γ组为 10 9%。结论 IFN-γ可抑制动脉损伤后的 SMC增殖和迁移并减轻 TGF-β1的促 SMC增殖和迁移的作用。并且 ,IFN -γ可以抑制 SMC MMP-2活力?

Objective To investigate the role of interfer on-γ on smooth muscle cells proliferation and migration after balloon injury.Methods Animal model of rabbit iliac artery balloon inj ury was set up, smooth muscle cells derived from injured artery were cultured.Sm ooth muscle cells were divided into four groups (control, TGF-β 1, IFN-γ, T GF-β 1 and IFN-γ).Cells from each group were treated with medium or TGF-β 1 (10 μg/L) and/or IFN-γ(500 kU/L) for 72 h separately. Smooth muscle cell proliferation was determined by cell count and MTT, migration of smooth muscle c ells was also detected. Matrix metalloproteinase-2 was also detected by zymogr aphy.Results Our results showed that, compared with group co ntrol,TGF-β 1 increased cell count and migration distance(P<0.01), and cell proliferation inhibiting rate of MTT was -19.4%; whereas IFN-γ decrease d cell count and migration distance (P<0.01), and cell proliferation inhibit ing rate of MTT was 15.8%; TGF-β 1 combined with IFN-γ increased cell cou nt and migration distance(P<0.01),but the value was lower than that in cel ls treated by TGF-β 1, proliferation inhibiting rate was -9.1%. Zymography sh owed that MMP-2 could be detected in all groups, and pro-MMP-2 could also be detected in cells treated by TGF-β 1. Compared with group control, the relati ve activities of MMP-2 in cells treated with TGF-β 1, IFN-γ and TGF-β 1 together with IFN-γ were 143%,95%,and 109% repectively. Conclusions IFN-γ can inhibit smooth muscle cells pro liferation and migration and reduce the stimulation of TGF-β 1 on smooth musc le cells proliferation and migration. IFN-γ can also inhibit MMP-2 activity a nd lesson the stimulation of TGF-β 1 on MMP-2 activity.