Background The key premise of genetic research and treatment is to select the desired gene vector,ultrasound microbubbles as a new type of gene vector can safe,fast and effectively enhance the gene transfection and expression by reversibility increasing the permeability of cells. Objective This study was to observe the effects of ciliary neurotrophic factor(CNTF)gene mediated by ultrasound microbubbles intraocular transfer on visual function and retinal ganglion cell(BGCs)after optic nerve injury. Methods Fifty-five adult Sprague-Dawley(SD)rats were divided into 6 groups randomly,including normal control group(n=5),sham injury group(n=10),simple injury group(n=10),naked plasmid group(n=10),plasmid+ultrasonic irradiation group(n=10)and ultrasound microbubbles group(n=10).The model of optic nerve injury Was made by forceps clip on the fight optic nerve.and the corresponding intervene was performed in different groups.Flash visual evoked potential(F%背景选择理想的基因载体是当前基因研究和治疗的前提与关键,超声微泡造影剂作为一种新型的基因载体,可安全、快速、有效地增强目的基因的转染和表达.目的观察超声微泡造影剂介导睫状神经营养因子(CNTF)基因转染视神经损伤大鼠对视功能及视网膜神经节细胞(RGCs)存活的影响.方法SD大鼠60只随机分为正常对照组、假伤组、单纯损伤组、单纯质粒组、质粒+超声组、超声微泡组.采用钳夹大鼠右眼视神经法制作视神经损伤大鼠模型,然后各处理组大鼠分别接受相应的干预处理.质粒采用玻璃体腔注射法注入,超声则采用辐照法进行干预.造模前1 d和损伤后第7天检测每组大鼠闪光视觉诱发电位(F-VEP),并于第7天处死各组大鼠.应用荧光金逆行标记法计数各组大鼠RGCs存活数,采用实时定量聚合酶链反应(real-time PCR)检测大鼠视网膜中CNTF mRNA的表达量.结果损伤后第7天,单纯损伤组大鼠F-VEP P1波的隐含时较单纯质粒组、质粒+超声组和超声微泡组明显延长,超声微泡组P1波的隐含时短于单纯质粒组和质粒+超声组,差异均有统计学意义(P＜0.05).单纯质粒组、质粒+超声组和超声微泡组F-VEPP1波的振幅均高于单纯损伤组,超声微泡组高于单纯质粒组和质粒+超声组,差异均有统计学意义(P＜0.05).超声微泡组大鼠与正常对照组比较P1波振幅降低,差异有统计学意义(P＜0.05).单纯质粒组、质粒+超声组和超声微泡组平均RGCs数目均明显高于单纯损伤组,超声微泡组平均RGCs数多于单纯质粒组和质粒+超声组,但少于对照组及假伤组,差异均有统计学意义(P＜0.05).超声微泡组CNTF mRNA表达量高于正常对照组、假伤组、单纯损伤组、单纯质粒组和质粒+超声组,差异均有统计学意义(P＜0.05).结论超声微泡能增强CNTF基因在眼内的转染及表达,对视神经损伤大鼠RGCs早期有明显的保护作用,可有效促进视功能的恢复.
展开▼
