CAJ | 학술논문

目的探讨醛固酮对糖尿病肾病(DN)大鼠肾小管间质转分化的影响.方法采用Wistar大鼠腹腔注射链脲菌素(STZ,60 mg/kg)制备糖尿病模型,4周后尿蛋白＞30 mg/d为DN模型成功(n=16),随机分为DN组(n=8)和螺内酯组(SP组,n=8),以另8只正常大鼠作为对照组(N组).sP组给予螺内酯40 mg·kg-1·d-1,N组、DN组每日以等量清水灌胃.8周后处死大鼠,收集尿、血浆、肾组织检测24 h尿蛋白定量、血肌酐和肾脏病理变化;用放射免疫法检测血浆、肾组织醛固酮浓度;用免疫组化、Western印迹方法检测E钙黏蛋白(E-cadherin)、α平滑肌肌动蛋白(α-SMA)蛋白的表达;用RT-PCR的方法检测E-cadherin、α-SMA mRNA的表达.结果与对照组比较,DN组尿蛋白排泄量、血肌酐均显著增加(均P＜0.01),肾组织E-cadherin蛋白和mRNA表达显著下调(均P＜0.01),α-SMA蛋白和mRNA表达均显著上调(均P＜0.01).DN组大鼠肾组织醛固酮显著升高[(24.71±5.30)ng/g比(16.38±2.85)ng/g,P＜0.01],与尿蛋白排泄量、血肌酐、α-SMA蛋白表达呈正相关(r=0.737、0.574、0.688,均P＜0.05),与E-cadherin蛋白表达呈负相关(r=-0.659,P＜0.01).各组间血清醛固酮含量差异无统计学意义.与DN组比较,SP组大鼠尿蛋白排泄和血肌酐显著下降(均P＜0.01),E-cadherin蛋白和mRNA表达上调(均P＜0.05),而α-SMA蛋白和mRNA表达显著下调(均P＜0.01).结论 DN大鼠肾组织局部醛固酮参与了糖尿病肾病肾间质转分化,螺内酯可以阻断醛固酮与其受体结合,抑制肾小管间质转分化,从而起到肾脏保护作用.
목적 탐토철고동대당뇨병신병(DN)대서신소관간질전분화적영향.방법 채용Wistar대서복강주사련뇨균소(STZ,60 mg/kg)제비당뇨병모형,4주후뇨단백＞30 mg/d위DN모형성공(n=16),수궤분위DN조(n=8)화라내지조(SP조,n=8),이령8지정상대서작위대조조(N조).sP조급여라내지40 mg·kg-1·d-1,N조、DN조매일이등량청수관위.8주후처사대서,수집뇨、혈장、신조직검측24 h뇨단백정량、혈기항화신장병리변화;용방사면역법검측혈장、신조직철고동농도;용면역조화、Western인적방법검측E개점단백(E-cadherin)、α평활기기동단백(α-SMA)단백적표체;용RT-PCR적방법검측E-cadherin、α-SMA mRNA적표체.결과 여대조조비교,DN조뇨단백배설량、혈기항균현저증가(균P＜0.01),신조직E-cadherin단백화mRNA표체현저하조(균P＜0.01),α-SMA단백화mRNA표체균현저상조(균P＜0.01).DN조대서신조직철고동현저승고[(24.71±5.30)ng/g비(16.38±2.85)ng/g,P＜0.01],여뇨단백배설량、혈기항、α-SMA단백표체정정상관(r=0.737、0.574、0.688,균P＜0.05),여E-cadherin단백표체정부상관(r=-0.659,P＜0.01).각조간혈청철고동함량차이무통계학의의.여DN조비교,SP조대서뇨단백배설화혈기항현저하강(균P＜0.01),E-cadherin단백화mRNA표체상조(균P＜0.05),이α-SMA단백화mRNA표체현저하조(균P＜0.01).결론 DN대서신조직국부철고동삼여료당뇨병신병신간질전분화,라내지가이조단철고동여기수체결합,억제신소관간질전분화,종이기도신장보호작용.
Objective To explore the effect of aldosterone on renal epithelialmesenchymal transition in streptozocin(STZ)-induced diabetic nephropathy rats. Methods Wistar rats were intraperitoneally injected with STZ(60 mg/kg)for the preparation of diabetic model.After 4 weeks,the rats with urinary protein＞30 mg/d were regarded as successful diabetic nephropathy(n=16),and were randomly divided into diabetic nephropathy(DN group,n=8)and spironolactone group(SP group,n=8).Then eight healthy rats were selected randomly as control group(N group,n=8).SP group rats were treated with spironolactone 40 mg·kg-1·d-1,and N group and DN group rats were given equal water.After 8 weeks,rats were sacrificed to collect urine,blood plasma,kidney tissue for detection of 24 h urinary protein,creatinine and renal pathological changes.Aldosterone concentration in plasma and kidney tissue was detected by mdioimmunoassay;E-cadherin,α-SMA protein expression by immunohistochemistry,Western blotting; E-cadherin,α-SMA mRNA expression by RT-PCR. Results Compared with N group,serum creatinine, urinary protein excretion in the DN rats were significantly higher (P＜0.01,respectively), E-cadhefin protein and mRNA were significantly reduced (P＜0.01, respectively),α-SMA protein and mRNA expression was up-regulated (P＜0.01, respectively). Aldosterone level of kidney tissue in DN rats was increased obviously [(24.71±5.30) ng/g vs (16.38±2.85) ng/g, P＜0.01], which was positively correlated with urinary protein excretion, serum creatinine and α-SMA protein (r=0.737, 0.574, 0.688, P＜0.01, respectively), and negatively correlated with E-cadherin protein (r=-0.659, P＜0.O1). While no significant difference was found in serum aldosterone among three groups. Compared with DN rats, urinary protein excretion, serum creatinine were reduced (P＜0.01, respectively), E-cadherin protein and mRNA were increased (P＜0.01, respectively), α-SMA protein and mRNA expression were decreased (P ＜0.01, respectively) in SP group rats.Conclusions Local aldosterone involves in renal epithelial-mesenchymal transition in diabetic nephropathy rat. Spironolactone can block the effect of aldosterone and play a role in renal protection.