CAJ | 학술논문

目的探讨血管紧张素Ⅱ( AngⅡ)诱导下大鼠肾脏及条件永生性小鼠足细胞c-Abl的表达变化.方法采用AngⅡ泵(400ng·kg-1·min-1)植入SD大鼠皮下的方法建立AngⅡ输注模型,24只大鼠成模后被随机分为AngⅡ输注2周组、AngⅡ输注4周组、AngⅡ输注+替米沙坦(ARB,3 mg·kg-1·min-1)干预2周组及4周组,同时设生理盐水输注组和健康对照组,每组6只.分别于成模后2周末、4周末处死大鼠取肾.电镜下观察肾脏足细胞超微结构的改变；免疫荧光法检测肾脏c-Abl表达；实时PCR及Western印迹检测c-Abl mRNA及蛋白水平的改变.对于体外培养条件永生性小鼠足细胞,免疫荧光法检测足细胞肾脏c-Abl的表达,实时PCR及Western印迹法检测足细胞在AngⅡ不同作用浓度(10-9 mol/L～10-6 mol/L)及不同作用时间点(0h、3h、6h、12 h、24 h)c-Abl mRNA和蛋白水平的变化.结果 (1)免疫荧光检测结果显示肾脏足细胞有c-Abl表达；实时PCR及Western印迹结果显示AngⅡ输注2周组和4周组大鼠肾脏c-Abl表达增加(P＜0.05),ARB干预组大鼠肾脏c-Abl表达较同期AngⅡ输注组均有降低(P＜0.05).(2)体外培养的条件永生性小鼠足细胞胞质及胞核有c-Abl表达.AngⅡ可诱导培养的小鼠足细胞c-Abl mRNA及蛋白表达增加(P＜0.05),且呈时间和剂量依赖性.结论 c-Abl在肾足细胞及体外培养足细胞均有表达,AngⅡ可诱导c-Abl表达上调.c-Abl可能参与了AngⅡ诱导的足细胞损伤.
목적 탐토혈관긴장소Ⅱ( AngⅡ)유도하대서신장급조건영생성소서족세포c-Abl적표체변화.방법 채용AngⅡ빙(400ng·kg-1·min-1)식입SD대서피하적방법건립AngⅡ수주모형,24지대서성모후피수궤분위AngⅡ수주2주조、AngⅡ수주4주조、AngⅡ수주+체미사탄(ARB,3 mg·kg-1·min-1)간예2주조급4주조,동시설생리염수수주조화건강대조조,매조6지.분별우성모후2주말、4주말처사대서취신.전경하관찰신장족세포초미결구적개변；면역형광법검측신장c-Abl표체；실시PCR급Western인적검측c-Abl mRNA급단백수평적개변.대우체외배양조건영생성소서족세포,면역형광법검측족세포신장c-Abl적표체,실시PCR급Western인적법검측족세포재AngⅡ불동작용농도(10-9 mol/L～10-6 mol/L)급불동작용시간점(0h、3h、6h、12 h、24 h)c-Abl mRNA화단백수평적변화.결과 (1)면역형광검측결과현시신장족세포유c-Abl표체；실시PCR급Western인적결과현시AngⅡ수주2주조화4주조대서신장c-Abl표체증가(P＜0.05),ARB간예조대서신장c-Abl표체교동기AngⅡ수주조균유강저(P＜0.05).(2)체외배양적조건영생성소서족세포포질급포핵유c-Abl표체.AngⅡ가유도배양적소서족세포c-Abl mRNA급단백표체증가(P＜0.05),차정시간화제량의뢰성.결론 c-Abl재신족세포급체외배양족세포균유표체,AngⅡ가유도c-Abl표체상조.c-Abl가능삼여료AngⅡ유도적족세포손상.
Objective To evaluate the effects of angiotensin Ⅱ (Ang Ⅱ )infusion on renal c-Abl expression in vivo,and on podocyte c-Abl expression change in cultured mouse podocytes.Methods Twenty four male Sprague-Dawley rats (Group C,D,E and F) were assigned to receive Ang Ⅱ (400 ng· kg-1 min-1) by osmotic minipump and of which 12 rats (Group D and F) were assigned to receive telmisartan (3 mg·kg-1·d-1),six rats received normal saline(Group B),and six rats were used as normal control (Group A).Animals were sacrificed at day 14 (Group C and D),day 28 (Group E and F) respectively.Conditionally immortalized mouse podocytes were used in vitro.Podocytes were studied 2 weeks after thermoswitching from 33℃ to 37℃.Cells were fetal bovine serum (FBS) starved for at least 12 hours prior to stimulation.The cultured podocytes were treated with Ang Ⅱ doses ranging from 10 -9 mol/L to 10 -6 mol/L and for different hours.Expression of renal and podocytes c-Abl was examined by immunofluorescence staining,real-time PCR and Western blotting. Results (1) Distribution of c-Abl expression was mainly in the cytoplasm and nuclear of the podocytes in vivo and in vitro. (2) Expressions of c-Abl mRNA and protein were increased in Ang Ⅱ-infused rat podocytes and Ang Ⅱ-induced cultured mouse podocytes (P＜0.05),and the effects of Ang Ⅱ were dose-dependent and time-dependent in vitro.Conclusion There are c-Abl mRNA and protein expression in podocytes,and c-Abl may play a critical role in the pathogenesis of Ang Ⅱ -induced podocyte injury.