福辛普利和缬沙坦抑制AngⅡ诱导的大鼠肾小管上皮细胞klotho基因表达下调  

Role of fosinopril and valsartan on inhibiting klotho gene down-expression induced by Ang Ⅱ

作　　者：林书典[1] 周巧玲[2] 詹锋[1] 

机构地区：[1]海南省人民医院肾病内科,海南海口570311 [2]中南大学湘雅医学院湘雅医院肾病内科,湖南长沙410008

出　　处：《基础医学与临床》2011年第1期25-31,共7页Basic and Clinical Medicine

基　　金：海南省自然科学基金(309087)

摘　　要：目的探讨福辛普利(Fos)和缬沙坦(Val)对血管紧张素Ⅱ(AngⅡ)抑制NRK-52E细胞klotho基因表达的干预作用及klotho与转化生长因子-β1(TGF-β1)的关系。方法将大鼠肾小管上皮细胞(NRK-52E)分为对照组、AngⅡ(10-7mol/L)组、AngⅡ分别再加Fos(10-5mol/L)组、Val(10-5mol/L)组和Fos+Val组,培养24 h后,用反转录-多聚酶链反应(RT-PCR)检测klotho和TGF-β1mRNA表达;用免疫组织化学法和Western blot检测klotho和TGF-β1蛋白表达。对klotho和TGF-β1蛋白表达进行直线相关分析。结果 (1)AngⅡ诱导NRK-52E中TGF-β1mRNA表达上调,同时使klothomRNA表达下调,经Fos或Val或Fos+Val干预后,TGF-β1mRNA表达明显受抑制,而klothomRNA显著上调表达(P<0.05)。(2)Fos、Val和Fos+Val均明显上调AngⅡ诱导的klotho蛋白低表达(P<0.05),同时抑制TGF-β1蛋白高表达(P<0.05)。(3)klotho和TGF-β1蛋白表达呈直线负相关(r=-0.717,P<0.05)。结论福辛普利和缬沙坦可上调AngⅡ诱导的klotho基因低表达,抑制TGF-β1高表达,klotho基因mRNA表达增强可能与TGF-β1表达下调有关。Objective To investigate role of fosinopril（Fos） and Valsartan（Val） on klotho gene expression induced by Ang Ⅱ and the relationship between klotho and TGF-β1.Methods Culture cells,NRK-52E,were incubated with Ang Ⅱ,Fos and Val.Five groups,control,Ang Ⅱ（10^-7mol/L）,Ang Ⅱ（10^-7mol/L）＋Fos（10^-5mol/L）,Ang Ⅱ（10^-7mol/L）＋Val（10^-5mol/L） and Ang Ⅱ（10^-7mol/L）＋Fos（10^-5mol/L）＋Val（10^-5mol/L）,were divided to observe the expression of the klotho and TGF-β1 mRNA and protein.The klotho and TGF-β1 mRNA were detected by reverse transcription-polymerase chain reaction（RT-PCR）,immunohistochemistry（IHC） and Western blot was applied to determine the klotho and TGF-β1 protein expression.The linear relationship between klotho and TGF-β1 protein expression was analyzed by Pearson＇s.Results（1）Ang Ⅱ induced anup-regulated expression of TGF-β1 mRNA,and associated with down-regulated klotho mRNA in NRK-52E as compared to control（all P〈0.05）;in cells incubated with Fos or Val or booth of all,the klotho mRNA expression was enheanced,and the TGF-β1 mRNA expression was inhibited;combined Fos and Val could not enhance the expression of klotho gene,no significant different was shown as compared to the Fos group and Val group.（2）IHC and Western blot revealed that klotho protein was increased,TGF-β1 protein was decreased in Fos group,Val group and combination group as compared to Ang Ⅱ group,but no significantly different among the three groups was shown as compared to the Val group or Fos group.（3）Negative relationship was showed between klotho and TGF-β1 protein expression（r=-0.717,P〈0.05）.Conclusion Fosinopril and valsartan can enhance the expression of klotho gene,and down-regulate TGF-β1 expression in NRK-52E,the increased level of klotho expression may be one of the important factor for preventing the renal fibrosis induced by TGF-β1.

关 键 词：福辛普利 缬沙坦 KLOTHO基因 生长转换因子-β1 

分 类 号：R965[医药卫生—药理学]