中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2013年
2期
114-118
,共5页
黄新忠%温东海%张敏%谢琼虹%马乐婷%任月衡%关熠%陈靖%林善锬
黃新忠%溫東海%張敏%謝瓊虹%馬樂婷%任月衡%關熠%陳靖%林善錟
황신충%온동해%장민%사경홍%마악정%임월형%관습%진정%림선담
肾切除术%转化生长因子β%Smad蛋白质类%Sirt1蛋白%白藜芦醇
腎切除術%轉化生長因子β%Smad蛋白質類%Sirt1蛋白%白藜蘆醇
신절제술%전화생장인자β%Smad단백질류%Sirt1단백%백려호순
Nephrectomy%Transforming growth factor beta%Smad proteins%Sirt1 protein%Resveratrol
目的 探讨白藜芦醇(RSV)对5/6肾切除大鼠肾脏的保护作用及对转化生长因子(TGF)-β-Smad信号通路的影响.方法 将50只雄性SD大鼠随机分为3组:假手术组(Sham组,n=10)、5/6肾切除组(Nx组,n=20)和5/6肾切除+RSV治疗组(Nx+ RSV组,n=20),术后1周开始每天给予生理盐水或RSV(20 mg/kg)灌胃.建模后每4周收集24 h尿液检测尿蛋白含量,12周后处死大鼠,收集血和肾组织标本观察血肌酐和肾组织病理改变.免疫组化法观察各组大鼠肾组织TGF-β、结缔组织生长因子(CTGF)、纤连蛋白(FN)和Ⅰ型胶原(Col Ⅰ)变化.Western印迹法检测肾组织Smad3、磷酸化和乙酰化Smad3表达,免疫共沉淀检测Sirt1与Smad3的相互结合.结果 Nx组在术后12周尿蛋白量[(152.14±30.49) mg/24 h比(25.34±7.54) mg/24 h]、血肌酐[(111.60±21.50) μmol/L比(53.90± 11.59) μmol/L]和肾小球硬化指数(1.56±0.34比0.35±0.08)均显著高于Sham组(均P＜0.01),RSV治疗可显著抑制上述改变(均P＜0.01).免疫组化结果显示Nx组大鼠FN、Col Ⅰ、TGF-β和CTGF的表达高于Sham组(均P＜ 0.01),RSV治疗能显著抑制上述改变.Western印迹显示,Nx组大鼠磷酸化Smad3和乙酰化Smad3的表达显著高于Sham组(均P＜0.05),RSV治疗可显著降低乙酰化Smad3的表达(P＜0.05),但并未显著降低磷酸化Smad3的表达.免疫共沉淀研究显示Sirt1与Smad3相互作用.结论 RSV可改善肾切除大鼠蛋白尿、肾功能和纤维化等指标,其作用机制可能与激活Sirt1,降低乙酰化Smad3的表达有关,提示Sirt1可能是慢性肾脏病潜在的治疗靶点.
目的 探討白藜蘆醇(RSV)對5/6腎切除大鼠腎髒的保護作用及對轉化生長因子(TGF)-β-Smad信號通路的影響.方法 將50隻雄性SD大鼠隨機分為3組:假手術組(Sham組,n=10)、5/6腎切除組(Nx組,n=20)和5/6腎切除+RSV治療組(Nx+ RSV組,n=20),術後1週開始每天給予生理鹽水或RSV(20 mg/kg)灌胃.建模後每4週收集24 h尿液檢測尿蛋白含量,12週後處死大鼠,收集血和腎組織標本觀察血肌酐和腎組織病理改變.免疫組化法觀察各組大鼠腎組織TGF-β、結締組織生長因子(CTGF)、纖連蛋白(FN)和Ⅰ型膠原(Col Ⅰ)變化.Western印跡法檢測腎組織Smad3、燐痠化和乙酰化Smad3錶達,免疫共沉澱檢測Sirt1與Smad3的相互結閤.結果 Nx組在術後12週尿蛋白量[(152.14±30.49) mg/24 h比(25.34±7.54) mg/24 h]、血肌酐[(111.60±21.50) μmol/L比(53.90± 11.59) μmol/L]和腎小毬硬化指數(1.56±0.34比0.35±0.08)均顯著高于Sham組(均P＜0.01),RSV治療可顯著抑製上述改變(均P＜0.01).免疫組化結果顯示Nx組大鼠FN、Col Ⅰ、TGF-β和CTGF的錶達高于Sham組(均P＜ 0.01),RSV治療能顯著抑製上述改變.Western印跡顯示,Nx組大鼠燐痠化Smad3和乙酰化Smad3的錶達顯著高于Sham組(均P＜0.05),RSV治療可顯著降低乙酰化Smad3的錶達(P＜0.05),但併未顯著降低燐痠化Smad3的錶達.免疫共沉澱研究顯示Sirt1與Smad3相互作用.結論 RSV可改善腎切除大鼠蛋白尿、腎功能和纖維化等指標,其作用機製可能與激活Sirt1,降低乙酰化Smad3的錶達有關,提示Sirt1可能是慢性腎髒病潛在的治療靶點.
목적 탐토백려호순(RSV)대5/6신절제대서신장적보호작용급대전화생장인자(TGF)-β-Smad신호통로적영향.방법 장50지웅성SD대서수궤분위3조:가수술조(Sham조,n=10)、5/6신절제조(Nx조,n=20)화5/6신절제+RSV치료조(Nx+ RSV조,n=20),술후1주개시매천급여생리염수혹RSV(20 mg/kg)관위.건모후매4주수집24 h뇨액검측뇨단백함량,12주후처사대서,수집혈화신조직표본관찰혈기항화신조직병리개변.면역조화법관찰각조대서신조직TGF-β、결체조직생장인자(CTGF)、섬련단백(FN)화Ⅰ형효원(Col Ⅰ)변화.Western인적법검측신조직Smad3、린산화화을선화Smad3표체,면역공침정검측Sirt1여Smad3적상호결합.결과 Nx조재술후12주뇨단백량[(152.14±30.49) mg/24 h비(25.34±7.54) mg/24 h]、혈기항[(111.60±21.50) μmol/L비(53.90± 11.59) μmol/L]화신소구경화지수(1.56±0.34비0.35±0.08)균현저고우Sham조(균P＜0.01),RSV치료가현저억제상술개변(균P＜0.01).면역조화결과현시Nx조대서FN、Col Ⅰ、TGF-β화CTGF적표체고우Sham조(균P＜ 0.01),RSV치료능현저억제상술개변.Western인적현시,Nx조대서린산화Smad3화을선화Smad3적표체현저고우Sham조(균P＜0.05),RSV치료가현저강저을선화Smad3적표체(P＜0.05),단병미현저강저린산화Smad3적표체.면역공침정연구현시Sirt1여Smad3상호작용.결론 RSV가개선신절제대서단백뇨、신공능화섬유화등지표,기작용궤제가능여격활Sirt1,강저을선화Smad3적표체유관,제시Sirt1가능시만성신장병잠재적치료파점.
Objective To investigate the protective effect of resveratrol (RSV) on 5/6 nephrectomized rats and its mechanism.Methods Fifty male SD rats were randomly divided into three groups:sham operated (Sham,n =10),5/6 nephrectomy (Nx,n =20),and 5/6 nephrectomy + RSV 20 mg/kg (Nx + RSV,n =20).RSV or normal saline was administered one week after 5/6 nephrectomy.Proteinuria was detected every 4 weeks.Serum creatinine and the renal pathological changes were measured after 12 weeks.Immunohistochemisty staining of fibronectin (FN),collagen Ⅰ,transforming growth factor-β (TGF-β) and connective tissue growth factor (CTGF) were used to analyze the changes of renal fibrosis.Western blotting was used to measure the expression of Smad3,phospho-Smad3,and acety-Smad3.Immunoprecipitation was used to detect the interaction between Sirt1 and Smad3.Results Compared with the sham operated rats,subtotal nephrectomy significantly increased proteinuria [(152.14±30.49) mg/24 h vs (25.34±7.54) mg/24 h],serum creatinine[(111.60±21.50) μmol/L vs (53.90±11.59) μmol/L],glomerular sclerosis index (1.56±0.34 vs 0.35±0.08) and the expressions of fibronectin,collagen Ⅰ,TGF-β and CTGF in renal tissue at 12 weeks after operation (all P ＜ 0.01),and RSV treatment significantly inhibited the above up-regulations (all P ＜ 0.01).Compared with the sham operated rats,subtotal nephrectomy increased the expression of phospholylation and acetylation of Smad3.RSV treatment significantly reduced the expression of acety-Smad3,but had no effect on the phospho-Smad3.Immunoprecipitation revealed a binding effect of Smad3 with Sirt1.Conclusions RSV treatment can attenuate proteinuria,protect renal function and inhibit renal fibrosis in 5/6 nephrectomized rats.This renal protective effect is associated with reduced Smad3 acetylation and activation of Sirt1,which suggesting that Sirt1 may be a potential therapeutic target of CKD.
