PEP-1-SOD1融合蛋白抑制血管紧张素Ⅱ诱导的大鼠心脏成纤维细胞Ⅰ型胶原的合成  

Inhibitory effect of PEP-1-SOD1 fusion protein on synthesis of collagen Ⅰ induced by angiotensin Ⅱ in rat cardiac fibroblasts

作　　者：杨波[1] 谭利国[1,2] 张蕾[2] 郑飞[2] 孔霞[2] 郭凌郧[2] 杨建业[2] 王家宁[2] 

机构地区：[1]武汉大学人民医院心内科,湖北武汉430060 [2]湖北医药学院附属人民医院临床研究所,湖北十堰442000

出　　处：《新乡医学院学报》2013年第8期613-618,共6页Journal of Xinxiang Medical University

基　　金：湖北省高等学校优秀中青年团队计划项目(编号:T200811)

摘　　要：目的研究穿透肽(PEP-1)介导的铜-锌超氧化物歧化酶1(SOD1)(PEP-1-SOD1)融合蛋白对血管紧张素Ⅱ(ANGⅡ)诱导的大鼠心脏成纤维细胞(CFbs)Ⅰ型胶原合成的影响,并探讨其机制。方法利用基因工程技术表达和纯化PEP-1-SOD1融合蛋白。提取原代大鼠CFbs并传代培养,采用2～5代细胞进行实验,实验分为对照组、ANGⅡ诱导组、SOD1预处理组和PEP-1-SOD1预处理组。SOD1预处理组和PEP-1-SOD1预处理组细胞分别以2μmol.L-1的SOD1和PEP-1-SOD1预处理2 h后,再给1μmol.L-1的ANGⅡ诱导24 h,用免疫荧光法检测细胞内超氧阴离子(O2-)水平,微量丙二醛(MDA)检测试剂盒检测各组细胞MDA含量。结果 Western blot和细胞免疫荧光结果显示PEP-1-SOD1成功穿透入CFbs;Western blot结果显示,与ANGⅡ诱导组比较,PEP-1-SOD1预处理组Ⅰ型胶原和α平滑肌肌动蛋白表达显著降低(P<0.01),细胞内O2-和MDA水平也显著降低(P<0.01)。结论 PEP-1-SOD1融合蛋白穿透入大鼠心脏CFbs内,通过降低细胞内O2-水平,抑制CFbs的激活,减少Ⅰ型胶原的合成。Objective To investigate the effect of PEP-1-SOD1 fusion protein on synthesis of collagen Ⅰ of rats induced by angiotensin Ⅱ（ANG Ⅱ） in rat cardiac fibroblasts（CFbs） and discuss the mechanism.Methods The genetic engineering technique was used to express and purify PEP-1-SOD1 fusion protein.Rat CFbs obtained from original generation of rats were cultured and passaged,and the cells from passage 2 to passage 5 were divided into four groups：control group,ANG Ⅱ induction group,SOD1 treatment group and PEP-1-SOD1 treatment group.The last two groups were pretreated with 2 μmol·L-1 SOD1 and PEP-1-SOD1 for 2 hours respectively,and induced by 1 μmol·L-1 angiotensin Ⅱ for 24 hours.And the levels of maleic dialdehyde（MDA） and O2-in cells were assayed with minimal MDA assay-kit and immunocytofluorescence,respectively.Results CFbs were penetrated successfully by PEP-1-SOD1.Compared with those in ANG Ⅱ induction group,the contents of collagen Ⅰ and alpha-smooth muscle actin reduced significantly（P 0.01）,and the levels of MDA and O2-in cells also decreased significantly in PEP-1-SOD1 treatment group（P 0.01）.Conclusion PEP-1-SOD1 fusion protein penetrated into rat CFbs suppresses activation of CFbs and lessens synthesis of collagen Ⅰ by decreasing the levels of O2-in cells.

关 键 词：穿透肽 铜-锌超氧化物歧化酶1 心脏成纤维细胞 血管紧张素Ⅱ Ⅰ型胶原 

分 类 号：R363[医药卫生—病理学]