南方医科大学学报

南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (11): 1654-.

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阿米洛利抑制尿激酶受体表达及减少蛋白尿的机制

何朝生，章斌，谢少庭，杨芸，马娟，史伟   

  • 出版日期:2014-11-20 发布日期:2014-11-20

Amiloride reduces proteinuria and inhibits podocyte uPAR in the 5/6 nephrectomy rats

  • Online:2014-11-20 Published:2014-11-20

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摘要： 目的观察阿米洛利在经典的5/6肾切除大鼠模型中对蛋白尿的影响，探讨其影响蛋白尿的机制。方法构建经典的5/6肾
切除大鼠模型，实验分为3组：假手术对照组（Sham）、5/6肾切除组（NTX）、5/6肾切除+阿米洛利干预组（NTX+阿米洛利）。采用
考马斯亮蓝法检测各组24 h 尿蛋白，利用免疫荧光共聚焦、实时荧光定量PCR 等技术，检测足细胞尿激酶受体（uPAR）蛋白和
uPAR mRNA 的表达情况。结果第2周时，与假手术对照组（Sham）相比较，5/6肾切除组（NTX）24 h尿蛋白明显升高（47.50±
28.05 mg vs 14.28±3.8 mg，P=0.023），有统计学意义；与NTX组相比，NTX+阿米洛利24 h尿蛋白无统计学意义（51.56±21.03 mg
vs 47.50±28.05 mg，P=0.748）。第12 周时，与NTX组（188.31±29.82）mg相比，NTX+阿米洛利组（121.37±31.14）mg、Sham组
（21.32±8.59）mg 24 h 尿蛋白增加量均进一步减少，具有统计学意义［P值分别为0.001，0.000］。激光共聚焦显微镜观察，与
Sham 组相比较，NTX组PLAUR mRNA表达明显增高（9.74±1.44 vs 1.01±0.13，P<0.01），有统计学意义；与NTX组相比较，
NTX+阿米洛利mRNA表达明显降低（9.74±1.44 vs 5.01±1.36，P<0.01），有统计学意义。结论阿米洛利可能通过抑制受损足细
胞uPAR 的表达，从而起到降蛋白尿、延缓肾小球硬化的作用。

Abstract: Objective To observe the effect of amiloride on the proteinuria of the 5/6 nephrectomy rats. Methods To establish the
5/6 nephrectomy rats model and divide the experiment into 3 groups, sham operated group(Sham), 5/6 nephrectomy model
group(NTX) and 5/6 nephrectomy with amiloride-treated group (NTX + amiloride, n=15). The concentration of protein and
mRNA of uPAR and the change of podocytes motility were detected by coomassiebluestaining, immunofluorence method and
real-time PCR. Results At second week, compared with Control group, the 24 h urine protein of NTX group was significantly
increased (47.50 ± 28.05 mg vs 14.28 ± 3.8 mg, P=0.023). There was no statistical significance in 24-hour urine protein between
NTX+amiloride group and NTX group (51.56±21.03 mg vs 47.50±28.05 mg, P=0.748). The same situation was also observed at
the time point of 12 week, comparing with NTX group, 24-hour urine protein decreased in Sham group (188.31±29.82 mg vs
21.32±8.59 mg, P=0.000) and NTX+amiloride group (188.31±29.82 mg vs 121.37±31.14 mg, P=0.000), with statistical significance
when comparing with Sham group, the expression of uPAR mRNA in NTX group was significantly increased (9.74 ± 1.44 vs
1.01±0.13, P=0.000). In contrast, the expression of uPAR mRNA in NTX rats treated with amiloride was significantly lower than
in NTX group (9.74±1.44 vs 5.01±1.36, P=0.000). Conclusion Amiloride can reduce the proteinuria of the 5/6 nephrectomy rats
model of transient proteinuria by inhibiting the induction of uPAR expression.