PEP-1-SOD1融合蛋白转导入大鼠肝脏组织的能力及酶活性  

TRANSDUCTION OF PEP-1-SOD1 FUSION PROTEIN INTO RAT LIVER TISSUES AND ENZYME ACTIVITY

作　　者：张友恩[1] 王家宁[1] 唐俊明[1] 杨建业[1] 郭凌郧[1] 黄永章[1] 付守芝[1] 

机构地区：[1]郧阳医学院附属人民医院临床医学研究所,十堰442000

出　　处：《肝胆外科杂志》2008年第6期460-463,共4页Journal of Hepatobiliary Surgery

基　　金：湖北省高等学校优秀中青年科技创新团队计划(T200811);十堰市重大科技项目(2006030Z6)

摘　　要：目的研究PEP-1-SOD1融合蛋白转导入大鼠肝脏组织的能力及其酶活性。方法实验分为SOD1组和PEP1-SOD1组,分别经尾静脉注射500μgSOD1和500μgPEP-1-SOD1融合蛋白入大鼠体内,于0.5,1,2,4,8和24h时间点取肝脏(n=6,每组,每个时间点)。免疫荧光技术检测PEP-1-SOD1的转导能力。黄嘌呤氧化酶法测定肝脏组织SOD1活性。结果SOD1蛋白不能进入肝脏组织内。PEP-1-SOD1可转导入肝脏组织内,SOD1活性于注射后0.5h开始升高,1h达高峰,持续存在达24h,两组各个时间点比较,差异有统计学意义(P<0.01)。SOD1组内各时间点比较差异无统计学意义(P>0.05)。结论PEP-1-SOD1以天然酶活性形式转导入肝脏组织,为进一步的动物模型实验及临床疾病进行蛋白质治疗提供理论基础。Objective To investigate the in vivo transduction capability of PEP-I-SOD1 fusion protein into rat liver tissues, and the SOD1 enzyme activity was measured. Methods The study included two groups： SODl-treated group and PEP-1-SODl-treated group. 500 μg of purified SOD1 and 500μg of purified PEP-1-SOD1 fusion proteins were administered in rats by tail vein, respectively. The livers were removed at designated times （0. 5, 1,2, 4, 8 and 24 h） after protein injection （n=6 for each group at each time point）. The transduction ability of transduced PEP-1-SOD1 fusion protein was detected with immuuofluorescecnce method. The liver SOD1 activity was measured with xanthine oxidase protocol. Results SOD1 alone could not be transduced into liver tissues, whereas PEP-1-SOD1 fusion protein could be transduced into liver tissues after 30 min via vein injection. The liver SOD1 activity of PEP-1- SODl-treated group was increased gradually after 30 min, and reached the peak at 1 h, then decreased gradually between 2～24 h. Conclusion PEP-I-SOD1 fusion protein can be transduced rapidly and efficiently into liver tissues in a native protein structure in vivo. These results open new possibilities for direct delivery of proteins into patients in the context of protein therapy, as well as for epigenetic experimentation with model organisms.

关 键 词：细胞穿透肽 PEP-1肽 肝 缺血再灌注损伤 铜 锌-超氧化物歧化酶 

分 类 号：R657[医药卫生—外科学]