rAAV介导hBDNF基因转染对急性高眼压兔眼视网膜BDNF表达的影响

目的 通过观察视网膜内源性脑源性神经营养因:F(BDNF)表达的变化,探讨玻璃体注射携带人脑源性神经营养因子(hBDNF)的重组腺伴随病毒(rAAV-BDNF)对急性高眼压兔眼神经损害的保护机制.方法 24只健康日本大耳白兔任选一眼作为造模眼(为模型组,共24眼),用生理盐水前房灌注法造成急性高眼压模型,对侧眼不作任何处理作为正常对照组(24眼).另24只健康日本大耳白兔任选一眼作为造模眼(为BDNF组,共24眼),BDNF组在造模前3d玻璃体内注射10μl rAAV-BDNF.于造模后第1、3、7、14d三组各摘除6只观察眼做病理切片,进行免疫组化染色,观察视网膜内源性BDNF表达.结果 模型组兔眼视网膜内源性BDNF表达阳性细胞数减少,BDNF组兔眼视网膜内源性BDNF表达阳性细胞数较对照组及模型组增加(P<0.05,P<0.01).结论 rAAV-BDNF基因转染通过增加视网膜内源性BDNF的表达起到神经保护作用.玻璃体注射是rAAV-BDNF转染视网膜的有效途径.

Abstract：

Objective To observe the changes in the expression of brain derived neurotrophic factor (BDNF) gene in the retina of rabbits with acute high intraocular pressure (IOP) after injection of recombinant adeno-associated virus (rAAV) vector containing human BDNF gene (rAAV-hBDNF), and investigate the neuroprotective mechanism of rAAV-hBDNF. Methods The unilateral eyes of 24 white rabbits were randomly chosen as the model group with high IOP induced by saline perfusion into the anterior chamber, and the contralateral eyes served as the control group without treatment. In another 24 white rabbits, 10 μl rAAV-BDNF was injected into the vitreous body of one of the eyes 3 days before induction of high IOP. On days 1,3,7, and 14 after perfusion, the bilateral eyes of 6 rabbits were excised for immunohistochemistry for the expression of endogenous BDNF gene in the retina. Results The number of BDNF-positive cells in the retina decreased after induction of high IOP, and injection of rAAV-hBDNF resulted in a significant increase in BDNF-positive cells as compared with the positive cell number in the high IOP model and control groups (P<0.05, P<0.01). Conclusion rAAV-mediated BDNF gene transfection can increase endogenous BDNF expression in the retina of rabbits with acute high IOP. Intravitreous injection is an effective pathway for rAAV-hBDNF gene transfection into the retina.

Effects of rAAV-mediated rhBDNF gene transfection on BDNF gene expression in the retina of a rabbit model of acute high intraocular pressure

Objective To observe the changes in the expression of brain derived neurotrophic factor (BDNF) gene in the retina of rabbits with acute high intraocular pressure (IOP) after injection of recombinant adeno-associated virus (rAAV) vector containing human BDNF gene (rAAV-hBDNF), and investigate the neuroprotective mechanism of rAAV-hBDNF. Methods The unilateral eyes of 24 white rabbits were randomly chosen as the model group with high IOP induced by saline perfusion into the anterior chamber, and the contralateral eyes served as the control group without treatment. In another 24 white rabbits, 10 μl rAAV-BDNF was injected into the vitreous body of one of the eyes 3 days before induction of high IOP. On days 1,3,7, and 14 after perfusion, the bilateral eyes of 6 rabbits were excised for immunohistochemistry for the expression of endogenous BDNF gene in the retina. Results The number of BDNF-positive cells in the retina decreased after induction of high IOP, and injection of rAAV-hBDNF resulted in a significant increase in BDNF-positive cells as compared with the positive cell number in the high IOP model and control groups (P<0.05, P<0.01). Conclusion rAAV-mediated BDNF gene transfection can increase endogenous BDNF expression in the retina of rabbits with acute high IOP. Intravitreous injection is an effective pathway for rAAV-hBDNF gene transfection into the retina.