PEP-1介导铜锌超氧化物歧化酶在小鼠顶叶皮质的转导  

Transductive ability of PEP-1-SOD1 into parietal cortex in mice

作　　者：董敏[1] 席刚明[2] 吴文春[1] 曹雅军[1] 李红云[1] 

机构地区：[1]湖北省襄樊市中医院,湖北襄樊441000 [2]湖北省郧阳医学院附属人民医院,湖北十堰442000

出　　处：《临床军医杂志》2010年第5期693-695,883,共4页Clinical Journal of Medical Officers

基　　金：湖北省高等学校科学研究青年发展项目(98B042)

摘　　要：目的研究细胞穿透肽(cell penetrating peptide,CPP)PEP-1介导铜,锌超氧化物歧化酶(Cu,Zn-superoxide dismutase,SOD1)在小鼠顶叶皮质的转导。方法将150只健康成年昆明小鼠随机分为PEP-1-SOD1组以及SOD1组,分别腹腔注射200μg PEP-1-SOD1或者SOD1蛋白,于给药后1、2、4、8 h以及24 h等5个不同时间点取顶叶皮质行W estern b lotting免疫荧光组化染色以及SOD1活性检测。结果 PEP-1-SOD1组给药后小鼠顶叶皮质出现外源性SOD1蛋白条带,显微镜下观察到转导阳性细胞,并且SOD1活性升高,以上指标均在4h时达到最高峰;SOD1组顶叶皮质任何时间点均未检测到目的蛋白条带,显微镜下未发现转导阳性细胞,皮质SOD1活性无明显变化。结论 PEP-1可以有效介导SOD1以天然活性形式转导进入小鼠顶叶皮质。Objective To explore the transductive ability of PEP-1-Cu,Zn-superoxide dismutase(PEP-1-SOD1) into parietal cortex in mice.Methods Totally 150 healthy adult male Kuming mice were categorized randomly into two groups,which were intraperitoneally injected 200 μg PEP-1-SOD1(PEP-1-SOD1 group) and 200 μg SOD1(SOD1 group),respectively.The animals were sacrificed for Western blotting,immunofluorescence staining and SOD1 activity at designed time points(1-,2-,4-,8-and 24 h after administration).Results Western blotting demonstrated that PEP-1-SOD1 was expressed in parietal cortex 1 h after injection and reached the peak at 4 h in PEP-1-SOD1 group;immunofluorescence indicated that the parietal cortex treated with PEP-1-SOD1 showed bright green fluorescent signals 1 h after injection and the fluorescent intensity reached the peak at 4 h.SOD1 activity of PEP-1-SOD1 group was increased gradually 1 h after injection and showed the peak at 4 h,and then decreased gradually within 24 h.Whereas the band of protein was not found in SOD1 group,there was no green fluorescent signal in the parietal cortex,and the SOD1 activity of SOD1 group maintained unchanged at all time points.Conclusion Native PEP-1-SOD1 transduction into murine cerebral tissue provides a basis for therapy of various disorders related to oxidative stress in central nerve system.

关 键 词：介导 铜锌超氧化物歧化酶 昆明小鼠 顶叶皮质 阳性细胞 显微镜下 天然活性 时间点 蛋白 细胞穿透肽 组化染色 条带 免疫荧光 镜下观察 活性检测 给药 腹腔注射 Western 外源性 指标 

分 类 号：R82[医药卫生—临床医学]