Experimental study of TGF-β2 antisense oligodeo- xynucleotide as an anti-scarring agent in glaucoma surgery

目的:观察TGF-β2反义寡核苷酸对青光眼滤过术后结膜囊成纤维细胞的转化、增生的影响.方法:制备兔青光眼滤过手术模型.右眼结膜下注射TGF-β2反义寡核苷酸(A组);左眼分别注射TGF-β2错义寡核苷酸(B组)、PBS(C组),于造模后4,7,14,28 d取术区结膜囊标本进行免疫组织化学染色和电镜观察.结果:A组眼压术后14 d和21 d比B组(P＜0.01)和C组(P＜0.05)显著降低;滤泡平均生存时间A组(17.2 d)较B组(14.5 d)、C组13.5 d)明显延长(P＜0.05);A组α-SMA和PCNA阳性成纤维细胞数明显少于B、C组;成纤维细胞的超微结构A组和B、C组比较无明显区别.结论:TGF-β2反义寡核苷酸可降低兔青光眼滤过术后眼压,延长滤泡生存时间,抑制兔青光眼滤过术后结膜囊成纤维细胞的转化、增生.AIM: Currently available anti-scarring regimens for glaucoma filtration surgery have potentially blinding complications and safer alternatives would be beneficial. This experiment is to investigate the effect of TGF-β2 antisense oligodeoxynucleotide on differentiation, proliferation of subconjunctival fibroblast following glaucoma filtration surgery.METHODS: Glaucoma filtration surgery were performed on both eyes of 28 rabbits. TGF-β2 antisense oligodeoxynudeotide was subconjunctivally injected in the right eyes (A group), and TGF-β2 missense oligodeoxynucleotide (B group)or PBS(C group) was used at the same method in the left eyes as controls. Rabbits were killed at 4,7,14 and 28 days after surgery. Intraocular pressure (IOP), bleb characteristics were recorded at different time point. Subconjunctival fibroblasts were examined by immunohistochemistry and electron microscopy.RESULTS: The IOP of rabbits in group A was significantly lower at 14 days (6.74± 1.18 mmHg) and 21 days (8.15± 1.97mmHg) after operation than the IOP in group B (8.53± 1.04,9.72± 1.09 mmHg)(P ＜0.01) and group C(8.79± 1.21, 9.43±1.27 mmHg) (P ＜0.05). The mean bleb survival time was longer (17.2 days) in group A than that of group B (14.5 days) and group C (13.5 days)(P＜0.05). The population of the cells expressing α -smooth muscle actin(α -SMA) and proliferating cell nuclear antigen (PCNA) was significantly reduced in group A compared with the group B and C. The ultrastructure of fibroblast was not altered by TGF-β2 anti-sense oligodeoxynucleotide.CONCLUSION:TGF-β2 antisense oligodeoxynucleotide can prevent the scar formation after glaucoma surgery by inhibit the differentiation and proliferation of subconjunctival fibroblast. It could be a potentially useful anti-scarring alternative for the prevention of late surgical failure.中国广东省深圳市科技局资助项目中国科技核心期刊(ISTIC)0110-14