TGF-β_1对大鼠高危角膜移植排斥反应及超微结构的影响  

Effects of TGF-β_1 on high risk keratoplasty rejection and ultrastructure in rats

作　　者：王炯[1] 周炼红[1] 邢怡桥[1] 谭锦泉[2] 黄晓丽[1] 吴丽[1] 

机构地区：[1]武汉大学人民医院眼科,湖北省武汉市430060 [2]武汉大学医学院免疫系,湖北省武汉市430072

出　　处：《眼科新进展》2010年第4期313-317,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30672290)~~

摘　　要：目的探讨重组腺相关病毒介导的转化生长因子-β1(recombinant adeno-associated virus mediated transforming growth factor-β,rAAV-TGF-β)抑制大鼠高危角膜移植术后免疫排斥反应的作用及对植片超微结构的影响。方法构建rAAV-TGF-β真核表达质粒载体。碱烧伤的方法建立角膜新生血管化动物模型,52只角膜新生血管化的SD大鼠接受穿透性角膜移植手术,分为2组,实验组(26只)Wistar大鼠角膜植片浸泡在含rAAV-TGF-β1(10×1012v.g..L-1)的DMEM/F12培养液中,37℃孵育30min,移植到SD大鼠角膜植床上;对照组(26只)Wistar大鼠角膜植片浸泡在DMEM/F12培养液中,37℃孵育30min,移植到SD大鼠角膜植床上。术后裂隙灯显微镜观察植片排斥情况,比较2组角膜植片的平均存活时间和各时间点排斥反应指数。术后9d、14d角膜植片行超微结构检查,免疫组织化学方法检测角膜组织中IFN-γ、TGF-β1的表达。结果对照组角膜植片平均存活时间为(10.75±1.91)d,实验组为(22.85±3.48)d,2组比较差异有显著统计学意义(P<0.01);实验组术后各时间点角膜植片排斥反应指数均明显低于对照组,差异均有显著统计学意义(均为P<0.01)。术后9d、14d角膜植片超微结构检查显示,实验组角膜植片基质层成纤维细胞凋亡与坏死明显减轻;免疫组织化学分析显示,实验组植片内IFN-γ表达低于对照组,TGF-β1表达高于对照组,差异均有统计学意义(P<0.05或0.01)。结论rAAV-TGF-β1能显著延长角膜植片存活时间,抑制大鼠高危角膜移植免疫排斥反应,改变植片的超微结构。Objective To investigate the effects of recombinant adeno-associated virus mediated transforming growth factor-β1（rAAV-TGF-β1）on high risk keratoplasty rejection and ultrastructure of corneal graft in rats.Methods The eukaryotic plasmid vector containing rAAV-TGF-β1 was constructed.Corneal neovascularization models were established by alkali burn.Fifty-two SD rats with Corneal neovascularization were operated with penetrating keratoplasty,and randomly divided into two groups.In experimental group（26 rats）,the donor corneal graft of Wistar rat was incubated in rAAV-TGF-β1（10×1012 v.g.·L-1）and Dulbecco＇s Modified Eagle Medium：Nutrient Mixture F-12（DMEM/F12）mixed culture medium for 30 minutes at 37℃ to be transplanted on SD rat.In control group（26 rats）,the donor graft of Wistar rat was treated in DMEM/F12 for 30 minutes at 37℃ to be transplanted on SD rats.Graft rejection was observed under slit lamp microscope.Mean survival time and rejection index of each stage were compared between the two groups.Ultrastructural changes of the corneal grafts were investigated,and the expression of IFN-γ and TGF-β1 was evaluated by immunohistochemistry at postoperative 9 days and 14 days.Results The mean survival time of control group was（10.75±1.91）days,but was（22.85±3.48）days in experimental group,there was significant difference between the two groups（P0.01）.Rejection index of experimental group was obviously lower than control group at different time points（All P0.01）.Stromal fibroblasts apoptosis and necrosis in corneal graft were obviously lightened in experimental group with examination of ultrastructure of corneal graft at postoperative 9 days and 14 days.In experimental group,the expression of TGF-β1 was higher and IFN-γ was lower than control group with immunohistochemical staining（P0.05 or 0.01）.Conclusion rAAV-TGF-β1 can significantly prolong the survival time of corneal graft,suppress the immunological rejection of high risk keratoplasty and change the ultr

关 键 词：重组腺相关病毒 高危角膜移植 免疫排斥反应 超微结构 转化生长因子-β1 干扰素-Γ 

分 类 号：R772.2[医药卫生—眼科]