摘要
目的:研究PEP-1介导铜,锌-超氧化物歧化酶(Cu,Zn-SOD1)转导入小鼠海马组织的能力及时间关系.方法:健康雄性昆明小鼠随机分为SOD1组及PEP-1-SOD1组,分别经腹腔注射SOD1蛋白及PEP-1-SOD1融合蛋白200μg,于注射后1,2,4,8,24h等5个时间点分别取小鼠海马组织进行检测(n=5).Western Blot测定PEP-1-SOD1蛋白的表达;免疫荧光组织化学方法测定PEP-1-SOD1在海马组织的分布;黄嘌呤氧化酶法测定SOD1活性.结果:PEP-1-SOD1组注射后1h,小鼠海马组织在Mr约26×103处出现目的蛋白条带,4h达高峰,24h仍有蛋白转导;SOD1组未发现目的蛋白.免疫荧光检测发现PEP-1-SOD1组海马组织中出现特异性绿色荧光;SOD1组未发现绿色荧光.PEP-1-SOD1组酶活性于注射后1h升高至(61.7±2.9)×103U/g,4h达峰值(82.6±4.2)×103U/g,之后开始下降,24h仍明显高于SOD1处理组(P<0.01);SOD1组酶活性各时间点相比较差异无统计学意义.结论:PEP-1可以介导SOD1以天然活性形式转导进入小鼠海马组织,4h达高峰,并保持活性至少达24h.
AIM： To explore the transductive ability of PEP-1- Cu, Zn-superoxide dismutase （PEP-1-SOD1） fusion protein into hippocampal tissue in mice and its persisting time. METHODS： Healthy adult male Kunming-mice were randomly divided into 2 groups： SOD1 treated group and PEP-1-SOD1 treated group. The mice were intraperitoneally injected with 200μg SOD1 in SOD1 treated group and 200 μg PEP-1-SOD1 fusion protein in PEP-1- SOD1 treated group respectively. The animals were sacrificed at 1, 2, 4, 8 and 24 h after injection （n = 5 ）. The animals＇ hippocampal specimens were examined with Western blot analysis, immunofluorescenee staining and SOD1 activity test. RESULTS： In PEP-1-SOD1 treated group, fusion protein was expressed with Mr of 26 × 10^3 1h after injection and reached the peak at 4 h after injection, while no protein expressed in SOD1 treated group. The hippocampal tissue treated with PEP-1-SOD1 fusion protein showed bright green fluorescent signal, but no green fluorescent signal was seen in SOD1 treated specimen. The SOD1 activity in PEP-1-SOD1 group increased to（61.7 ±2.9） ×10^3 U/g 1 h after injection, reached the peak of（82.6 ±4.2） ×10^3 U/g at 4 h after injection and then decreased gradually, but it was still higher than that in SOD1 treated group at 24 h after injection （P 〈 0.01 ）. CONCLUSION： PEP-1 mediates the transduction of natively active SOD1 into hippocampal region in mice and the enzyme activity can persist for no less than 24 h
出处
《第四军医大学学报》
北大核心
2009年第13期1184-1187,共4页
Journal of the Fourth Military Medical University
基金
湖北省高等学校科学研究青年发展项目(98B042)
