脂质体介导TFPI-2质粒DNA转染兔角膜基质细胞的实验研究  

Experimental Study of Plasmid TFPI-2 DNA Gene Transfection with Lipofectamine to Rabbit Corneal Keratocytes in Vitro

作　　者：袁静[1] 胡燕华[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科,湖北武汉430022

出　　处：《湖北民族学院学报（医学版）》2004年第1期5-8,共4页Journal of Hubei Minzu University(Medical Edition)

摘　　要：目的　探讨角膜基质细胞组织因子途径抑制物 2 (TFPI- 2 )的表达情况 ,为角膜新生血管的基因治疗提供方法。方法　体外兔角膜基质细胞原代、传代培养 ;脂质体介导人类TFPI- 2真核表达载体pBos -Cite -neo/TFPI- 2转染角膜基质细胞 ,RT -PCR ,Westernblot技术检测转染前后基质细胞中TFPI- 2mRNA及相应蛋白质的表达水平。结果　转染前后基质细胞TFPI- 2mRNA的表达水平IATFPI-2 /IAβ -actin分别为 0 .5 6± 0 .0 4、0 .78± 0 .0 3;蛋白质的表达水平IA值分别为 1 8.4± 1 .3、2 4 .2± 0 .5 ,差异均有显著意义。结论　培养角膜基质细胞可基础表达一定量的TF PI- 2 ,转染TFPI- 2质粒后使基质细胞TFPI- 2mRNA和蛋白质的表达增高 。Objective To observe the expression of TFPI-2 in corneal keratocytes in vitro and investigate the experimental basis for curing corneal neovascularization with gene-therapy.Methods Primary culture and subculture of rabbit keratocytes were established in vitro; Human TFPI-2 expression vector pBos-Cite-neo/TFPI-2 was transfected into keratocytes, transfected and nontransfected cells were screened for TFPI-2 mRNA and protein by reverse transcription-polymerase chain reaction and Western blot analysis, respectively. Results IA TFPI-2 /IA β-actin of TFPI-2 mRNA in transfected and nontransfected cells were 0.56±0.04、0.78±0.03 respectively ;IA of TFPI-2 protein were 18.4±1.3、24.2±0.5 respectively, there are significant difference between the two group cells (P<0.05).Conclusions TFPI-2 protein and gene expression level in transfected keratocytes were much higher than those of nontransfected keratocytes, which may offer the beneficial suggestions in the genetherapy.to corneal neovascularization.

关 键 词：脂质体介导 TFPI-2质粒 DNA转染 角膜基质细胞 实验研究 

分 类 号：R772.2[医药卫生—眼科]