含LacZ报告基因的pcDNA3.0裸质粒转染青光眼滤过术后家兔模型的表达  

Expression of naked plasmid pcDNA3.0 with LacZ gene in rabbit models after trabeculectomy

作　　者：喻箭[1] 李平[2] 周元国[2] 叶剑[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所眼科,中国重庆市400042 [2]第三军医大学大坪医院研究所分子生物学中心,中国重庆市400042

出　　处：《国际眼科杂志》2007年第3期658-660,共3页International Eye Science

摘　　要：目的:观察含LacZ报告基因的裸pcDNA3.0重组质粒在体转染小梁切除术后家兔眼的表达情况,建立动物模型,评价其应用价值,为利用裸质粒对抗青光眼术后瘢痕行基因治疗打下基础。方法:制作家兔双眼小梁切除术后模型,采用自身对照,术后1d在右眼术区局部注射含LacZ报告基因的pcD-NA30裸质粒0.1mL(100μg),左眼术区注射等剂量空质粒,通过X-gal染液原位染色检测注射后1,3,6,15d组织标本中报告基因LacZ的表达情况,绘制感染效率曲线。结果:重组pcDNA3.0裸质粒100μg可以转染术后滤过区结膜下成纤维细胞并表达携带的基因,表达高峰期3～6d,15d时仍可检测到少量表达细胞,对照侧各个时相点均未检测到表达。结论:重组pcDNA3.0裸质粒可以携带外源性基因转染家兔小梁切除术后术区成纤维细胞,适用于对青光眼滤过术后动物模型行基因治疗。AIM： To observe the expression of naked plasmid pcDNA3.0 with LacZ gene transferred to rabbits after trabeculectomy in vivo and to evaluate its application for the sake of treating scar after tTabeculectomy with naked plasmid. METHODS： The rabbits received bilateral filtering surgery. On postoprerative day 1, 0.1mL （100μg）naked plasmid pcD- A3.0 carrying report gene （LacZ.） was injected into the right eye filtering bleb and empty plasmid was injected into the left eye bleb as control of each rabbit. Then the rabbits were executed on the 1st, 3rd, 6th, 15th day respectively, enudeate their eyes and detect the transfection by histochemical assays of the tissue with X-gal. RESULTS： The dose （100μg） of naked plasmid pcDNA3.0 could transfer to fibroblasts underlying the conjunctiva and expressed report gene. The peak time of LacZ expression was in 3-6th days. On 15th day there also were a small quantity of transferred cells could be saw, but they were not expressed in the tissue of the control sides. CONCLUSION： The naked plasmid pcDNA3.0 can carry exogenic gene segment and expresses it in fibroblasts of filtering bleb of rabbit received trabeculectomy. The result indicates that this plasmid can be applied to transcjenic therapy of animal models after filtering surgery.

关 键 词：小梁切除术 基因治疗 裸质粒 LacZ报告基因 

分 类 号：R779.6[医药卫生—眼科]