中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2011年
9期
684-691
,共8页
刘军民%张欣洲%张万帆%贺晓蕾%戴勇%吴耀炯
劉軍民%張訢洲%張萬帆%賀曉蕾%戴勇%吳耀炯
류군민%장흔주%장만범%하효뢰%대용%오요형
再灌注损伤%肾切除术%干细胞%肾功能衰竭%肾脏修复%预后
再灌註損傷%腎切除術%榦細胞%腎功能衰竭%腎髒脩複%預後
재관주손상%신절제술%간세포%신공능쇠갈%신장수복%예후
Reperfusion injury%Nephrectomy%Stem cells%Kidney failure%Renal repair%Prognosis
目的 比较肾皮质大部分切除与肾缺血再灌注损伤对大鼠肾干、祖细胞的影响,探讨肾干、祖细胞在肾脏损伤修复中的意义及急性肾衰竭( AFR)和慢性肾衰竭(CRF)预后不同的可能机制.方法 肾动脉结扎再灌注和5/6肾皮质切除术(假手术为对照)分别制作SD大鼠ARF和CRF模型,定期监测血肌酐、尿素氮及24h尿蛋白量.于设定时间采集肾脏标本,HE染色检查病理改变,免疫荧光检测肾鲍曼囊区CD24、CD133及肾小球podocin表达；RT-PCR检测大鼠肾皮质区podocin mRNA表达和肾组织转化生长因子β1(TGF-β1)、Notch2、肝细胞生长因子(HGF)、成骨蛋白7(BMP7)和Pax-2 mRNA表达.分析5/6肾皮质切除术后Pax-2 mRNA表达量与podocin mRNA表达量及肾小球硬化指数(GSI)的相关性.结果 两种模型大鼠分别出现急、慢性肾衰竭的典型肾脏病理及功能变化.CRF组随时间延长肾小球硬化指数逐渐升高,于术后第14、30、60、90天分别为(2.34±0.28)％、( 25.12±5.67)％、(89.42±12.28)％和(171.23±32.28)％.与假手术组比较,ARF组不同时间点大鼠鲍曼囊区CD24+CD133+表达细胞分布无显著变化,而CRF组大鼠鲍曼囊区CD24+CD133+表达细胞逐渐减弱；ARF组肾小球podocin表达有短暂减少后迅速恢复,而CRF组肾小球podocin表达则进行性减少.与假手术组相比,ARF组HGF、BMP7mRNA表达升高(P＜0.05),而CRF组TGF-β1、Notch2 mRNA表达升高(P＜0.05),Pax-2和podocin mRNA表达均进行性减少(P＜0.05).后两者呈正相关(r=0.872),且均与GSI呈负相关(r=-0.906、-0.872,均P＜0.05).结论 肾脏缺血再灌注损伤对大鼠肾干、祖细胞无明显损伤,足细胞修复迅速,肾脏结构及功能完全恢复.肾皮质大部分切除引起大鼠肾干、祖细胞所处环境中生长抑制因子水平上调,促生长因子降低,导致肾干、祖细胞逐渐减损,足细胞修复缺陷,肾小球硬化及肾功能进行性衰竭.两种肾损伤对肾脏干、祖细胞的不同影响及由此产生肾脏再生修复功能的差异可能是其预后不同的主要机制.
目的 比較腎皮質大部分切除與腎缺血再灌註損傷對大鼠腎榦、祖細胞的影響,探討腎榦、祖細胞在腎髒損傷脩複中的意義及急性腎衰竭( AFR)和慢性腎衰竭(CRF)預後不同的可能機製.方法 腎動脈結扎再灌註和5/6腎皮質切除術(假手術為對照)分彆製作SD大鼠ARF和CRF模型,定期鑑測血肌酐、尿素氮及24h尿蛋白量.于設定時間採集腎髒標本,HE染色檢查病理改變,免疫熒光檢測腎鮑曼囊區CD24、CD133及腎小毬podocin錶達；RT-PCR檢測大鼠腎皮質區podocin mRNA錶達和腎組織轉化生長因子β1(TGF-β1)、Notch2、肝細胞生長因子(HGF)、成骨蛋白7(BMP7)和Pax-2 mRNA錶達.分析5/6腎皮質切除術後Pax-2 mRNA錶達量與podocin mRNA錶達量及腎小毬硬化指數(GSI)的相關性.結果 兩種模型大鼠分彆齣現急、慢性腎衰竭的典型腎髒病理及功能變化.CRF組隨時間延長腎小毬硬化指數逐漸升高,于術後第14、30、60、90天分彆為(2.34±0.28)％、( 25.12±5.67)％、(89.42±12.28)％和(171.23±32.28)％.與假手術組比較,ARF組不同時間點大鼠鮑曼囊區CD24+CD133+錶達細胞分佈無顯著變化,而CRF組大鼠鮑曼囊區CD24+CD133+錶達細胞逐漸減弱；ARF組腎小毬podocin錶達有短暫減少後迅速恢複,而CRF組腎小毬podocin錶達則進行性減少.與假手術組相比,ARF組HGF、BMP7mRNA錶達升高(P＜0.05),而CRF組TGF-β1、Notch2 mRNA錶達升高(P＜0.05),Pax-2和podocin mRNA錶達均進行性減少(P＜0.05).後兩者呈正相關(r=0.872),且均與GSI呈負相關(r=-0.906、-0.872,均P＜0.05).結論 腎髒缺血再灌註損傷對大鼠腎榦、祖細胞無明顯損傷,足細胞脩複迅速,腎髒結構及功能完全恢複.腎皮質大部分切除引起大鼠腎榦、祖細胞所處環境中生長抑製因子水平上調,促生長因子降低,導緻腎榦、祖細胞逐漸減損,足細胞脩複缺陷,腎小毬硬化及腎功能進行性衰竭.兩種腎損傷對腎髒榦、祖細胞的不同影響及由此產生腎髒再生脩複功能的差異可能是其預後不同的主要機製.
목적 비교신피질대부분절제여신결혈재관주손상대대서신간、조세포적영향,탐토신간、조세포재신장손상수복중적의의급급성신쇠갈( AFR)화만성신쇠갈(CRF)예후불동적가능궤제.방법 신동맥결찰재관주화5/6신피질절제술(가수술위대조)분별제작SD대서ARF화CRF모형,정기감측혈기항、뇨소담급24h뇨단백량.우설정시간채집신장표본,HE염색검사병리개변,면역형광검측신포만낭구CD24、CD133급신소구podocin표체；RT-PCR검측대서신피질구podocin mRNA표체화신조직전화생장인자β1(TGF-β1)、Notch2、간세포생장인자(HGF)、성골단백7(BMP7)화Pax-2 mRNA표체.분석5/6신피질절제술후Pax-2 mRNA표체량여podocin mRNA표체량급신소구경화지수(GSI)적상관성.결과 량충모형대서분별출현급、만성신쇠갈적전형신장병리급공능변화.CRF조수시간연장신소구경화지수축점승고,우술후제14、30、60、90천분별위(2.34±0.28)％、( 25.12±5.67)％、(89.42±12.28)％화(171.23±32.28)％.여가수술조비교,ARF조불동시간점대서포만낭구CD24+CD133+표체세포분포무현저변화,이CRF조대서포만낭구CD24+CD133+표체세포축점감약；ARF조신소구podocin표체유단잠감소후신속회복,이CRF조신소구podocin표체칙진행성감소.여가수술조상비,ARF조HGF、BMP7mRNA표체승고(P＜0.05),이CRF조TGF-β1、Notch2 mRNA표체승고(P＜0.05),Pax-2화podocin mRNA표체균진행성감소(P＜0.05).후량자정정상관(r=0.872),차균여GSI정부상관(r=-0.906、-0.872,균P＜0.05).결론 신장결혈재관주손상대대서신간、조세포무명현손상,족세포수복신속,신장결구급공능완전회복.신피질대부분절제인기대서신간、조세포소처배경중생장억제인자수평상조,촉생장인자강저,도치신간、조세포축점감손,족세포수복결함,신소구경화급신공능진행성쇠갈.량충신손상대신장간、조세포적불동영향급유차산생신장재생수복공능적차이가능시기예후불동적주요궤제.
Objective To compare the impacts of subtotal nephrectomy and ischemiareperfusion injury on renal stem cells and progenitor cells of rats,and to explore the significance of renal stem cells and progenitor cells for renal repair and the possible mechanisms of prognosis in rats with acute renal failure (ARF) or chronic renal failure (CRF).Methods Rats of CRF or ARF model underwent 5/6 nephrectomy or renal artery ligation and repedusion respectively,and rars in control group underwent sham operation.Scr,BUN and 24 hour urine protein were regularly measured.Kidney specimens were obtained at the set time for HE staining and fluorescence staining.Expressions of CD24,CD133 and podocin were detected by immunofluorescence.RT-PCR was performed to quantify the expression of transforming growth factor β1 (TGF-β1),Notch2,hepatocyte growth factor (HGF),bone morphogenetic protein 7 (BMP7) and Pax-2 mRNA in renal tissue and the expression of podocin mRNA in renal cortex.Correlation among the expressions of Pax-2 mRNA,podocin mRNA and glomemlosclerosis index were analyzed.Results The rats of two models presented typical ARF or CRF in renal pathology and function.Glomerulosclerosis index in CRF group increased gradually with time,which were (2.34±0.28)％,(25.12±5.67)％,(89.42±12.28)％ and (171.23±32.28)％ at day 14,day 30,day 60 and day 90 respectively.Compared with sham group,the CD24+CD133+ cells of the ARF rats showed no significant change in quantity and distribution,while the CRF rats showed gradual reduction of CD24 +CD133+ cells.The expression of podocin in glomerulus decreased temporarily and recovered finally after ischemiareperfusion injury,but decreased gradually after 5/6 nephrectomy.Compared with sham group,expression of TGF-β1,Notch2 mRNA in renal tissue was increased in CRF group,while the expression of HGF,BMP7 mRNA in renal tissue of ARF group were increased.Between the expression of Pax-2 mRNA in renal tissue and the expression of podocin mRNA in renal cortex,there was positive correlation in CRF group,while they both were negatively correlated with glomerulosclerosis index.Conclusions Ischemia-reperfusion injury makes no obvious impairment to renal progenitor cells.Having progressively injured the living environment of renal progenitor cells,subtotal nephrectomy reduces renal progenitor cells,and causes podocytes to repairing incompetently,which may be the main pathogenesis of CRF with poor prognosis.
