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目的探讨大鼠前房内注射阳离子聚合体/MY OC基因复合物转染小梁组织的有效性和安全性。设计实验研究。研究对象SD大鼠。方法用微量进样器将表达myocilin(MYOC)基因的重组质粒PCDNA3-MYOC与阳离子聚合体(PEI)的混合溶液20μl注射于SD大鼠前房内,于注射后不同时间点(1、3、7、14 d)摘除眼球,进行病理切片HE染色、荧光免疫组织化学染色以及透射电镜观察。主要指标病理切片、荧光免疫组织化学染色以及透射电镜观察小梁组织的变化。结果病理切片HE染色未见小梁组织明显炎症细胞浸润以及结构改变;荧光免疫组织化学染色显示注射后1 d可见到眼组织荧光着色;于转染后3 d小梁网荧光强度明显增强,至14 d荧光渐消失;透射电镜证实小梁细胞内可见PEI/DNA颗粒,溶酶体增多。结论前房内注射阳离子复合体介导的DNA质粒可将外源性基因安全有效地转染入小梁网组织。
Objective To investigate the efficacy and safety of intracameral injection of cationic polymer / MY OC gene into trabecular tissue in rats. Design experiment research. Study object SD rats. Methods Twenty microliters of the mixed solution of PCDNA3-MYOC and PEI, which express myocilin (MYOC) gene, was injected into the anterior chamber of SD rats with microinjector and injected at different time points after injection (1, 3, The eyeballs were excised for 7 and 14 days. The sections were stained with HE for immunohistochemical staining and transmission electron microscopy. The main indexes of pathological sections, fluorescence immunohistochemical staining and transmission electron microscopy trabecular tissue changes. Results There was no inflammatory cell infiltration and structural change in trabecular tissue observed by HE staining. The fluorescence staining of ocular tissue was observed on day 1 after injection. The fluorescence intensity of trabecular meshwork was significantly increased 3 days after transfection Fourteen days after the disappearance of fluorescence; transmission electron microscopy confirmed that trabecular cells PEI / DNA particles, lysosome increased. Conclusions Anterior chamber injection of cationic complex-mediated DNA plasmids can transfect exogenous genes safely and efficiently into the trabecular meshwork tissues.