促红细胞生成素对血管紧张素Ⅱ诱导的系膜细胞增殖及Ⅰ/Ⅳ型胶原和纤维连接蛋白表达的影响研究  

Effects of Erythropoietin on Ang Ⅱ-induced Mesangial Cell Proliferation and the Expression Levels of Col Ⅰ,Col Ⅳ and FN

作　　者：季芬[1,2] 徐玉音[1] 范亚平[1] 王锋[1] 杨斌[1] 

机构地区：[1]南通大学附属医院肾内科,江苏省南通市226001 [2]南通市第二人民医院肾内科

出　　处：《中国全科医学》2015年第26期3177-3181,共5页Chinese General Practice

基　　金：国家自然科学基金资助项目(81170689);南通市社会事业科技创新与示范计划项目(HS2011029)

摘　　要：目的探讨促红细胞生成素(EPO)对血管紧张素Ⅱ(AngⅡ)诱导的系膜细胞增殖及Ⅰ型胶原(ColⅠ)、Ⅳ型胶原(ColⅣ)和纤维连接蛋白(FN)表达的影响。方法将体外培养大鼠肾小球系膜细胞分为4组:对照组,AngⅡ组(0.1、1.0、10.0μmol/L),EPO组(1、10、100 U/L),AngⅡ+EPO组(EPO 1、10、100 U/L预处理1 h后加AngⅡ1.0μmol/L)。各组处理0、24、48 h时,CCK-8法检测系膜细胞增殖情况,免疫印迹法检测系膜细胞ColⅠ、ColⅣ和FN蛋白表达水平,ELISA法检测系膜细胞培养上清液中ColⅠ、ColⅣ和FN蛋白水平。结果单独给予AngⅡ或EPO均可刺激系膜细胞增殖,AngⅡ组亚组和EPO组亚组24、48 h时系膜细胞增殖水平均高于对照组,且48 h时高于24 h时,24 h时高于0 h时,差异均有统计学意义(P<0.05);单独给予AngⅡ或EPO均可促进系膜细胞合成及分泌ColⅠ、ColⅣ和FN,除EPO 1 U/L亚组ColⅠ蛋白表达水平与对照组间差异无统计学意义(P>0.05)外,AngⅡ组亚组和EPO组其他亚组ColⅠ、ColⅣ和FN蛋白表达水平均高于对照组,差异有统计学意义(P<0.05)。与AngⅡ1.0μmol/L+EPO 1 U/L亚组比较,AngⅡ1.0μmol/L+EPO 10 U/L亚组和AngⅡ1.0μmol/L+EPO 100 U/L亚组在24、48 h时系膜细胞增殖水平及ColⅠ、ColⅣ和FN蛋白表达水平均下降,差异有统计学意义(P<0.05),但与对照组及同浓度单纯EPO亚组比较,AngⅡ1.0μmol/L+EPO组各亚组24、48 h时系膜细胞增殖水平及ColⅠ、ColⅣ和FN蛋白表达水平均增高,差异有统计学意义(P<0.05)。结论 AngⅡ能刺激系膜细胞增殖,促进系膜细胞合成及分泌ColⅠ、ColⅣ和FN,EPO亦有一定程度类似作用,但两者同时存在时,EPO可拮抗AngⅡ诱导的系膜细胞增殖及ColⅠ、ColⅣ和FN的合成及分泌,提示EPO除可纠正肾性贫血外,可能对AngⅡ诱导的肾小球硬化具有抑制作用而保护肾功能。Objective To investigate the effects of erythropoietin on AngⅡ-induced mesangial cell proliferation and the expression levels of Col Ⅰ,Col Ⅳ and fibronectin（ FN）. Methods Rat mesangial cells were cultured in virto and were divided into 4 groups：control group,Ang Ⅱgroup（0. 1,1. 0,10. 0 mol/L）,EPO group（1,10,100 U/L）,and AngⅡ＋ EPO group（mesangial cells were pretreated with EPO 1,10,100 U/L for 1 h,then AngⅡ1. 0μmol/L was added）. After 0 h,24 h,48 h treatment,mesangial cell proliferation was assayed by CCK-8;the expression levels of ColⅠ,ColⅣand FN protein were detected by ELISA;the expression levels of Col Ⅰ,Col Ⅳ and FN in cell culture supernatant were detected by ELISA. Results Either AngⅡor EPO alone could stimulate mesangial cell proliferation;AngⅡgroup and EPO subgroups were higher than control group in cell proliferation levels at 24 h and 48 h,and the cell proliferation levels at 48 h were higher than 24 h,and those at 24 h were higher than 0 h（P〈0. 05）. Either Ang Ⅱ or EPO alone could stimulate the synthesis of mesangial＆amp;nbsp;cells and the secretion of Col Ⅰ, Col Ⅳ and FN;apart from EPO 1 U/L subgroup which was not significantly different from control group in the expression of ColⅠ（P＆gt;0. 05）,AngⅡ group and other EPO subgroups were all higher（P〈0. 05）than control group in the expression levels of ColⅠ,ColⅣand FN. Compared with AngⅡ1. 0 μmol/L＋EPO 1 U/L group,AngⅡ1. 0 μmol/L＋EPO 10 U/L group and AngⅡ1. 0 μmol/L＋EPO 100 U/L group were lower in 24 h and 48 h mesangial cell proliferation levels and the expression levels of ColⅠ,ColⅣand FN（P〈0. 05）,while compared with control group and EPO subgroups of the same concentrations,AngⅡ1. 0 μmol/L＋EPO（1,10,100 U/L）subgroups were higher in 24 h and 48 h mesangial cell proliferation levels and the expression levels of ColⅠ,Col Ⅳ and FN（P〈0. 05）. Conclusion Ang Ⅱ can increase the mesangial cell proliferation and promote the synthesis and secr

关 键 词：促红细胞生成素 血管紧张素Ⅱ 肾小球系膜细胞 胶原Ⅰ型 胶原Ⅳ型 纤连蛋白类 

分 类 号：R322.61[医药卫生—人体解剖和组织胚胎学]