| 周阳 闻萍 牛婧 徐静 丁昊 余燕婷 杨俊伟 江蕾.尿液外泌小体中miRNA的含量与肾纤维化的关系[J].,2016,16(33):6406-6409 |
| 尿液外泌小体中miRNA的含量与肾纤维化的关系 |
| miRNA Profiling in Urine Exosomes Indicates Renal TubulointerstitialFibrosis |
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| DOI： |
| 中文关键词: 外泌小体(exosomes) 微小RNA (miRNA) 肾纤维化 |
| 英文关键词: Exosomes miRNA Renal fibrosis |
| 基金项目:国家自然科学基金面上项目(81470971)；国家自然科学基金青年基金项目(31300955)；
江苏省自然科学基金面上项目(BK20141489) |
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| 中文摘要: |
| 目的：探讨尿液外泌小体(exosomes)中微小RNA(miRNA, miR)的变化与肾纤维化的关系，以寻找早期诊断肾纤维化的生物
标志物。方法：以行肾穿刺活检术并诊断为原发性肾脏病的患者为研究对象，其中，肾活检未发现肾纤维化的患者作为对照组，而
存在轻到中度肾纤维化的患者作为纤维化组。收集20 mL晨尿，用超速离心方法分离尿液exosomes，用电镜观察其形态，用定量
PCR方法检测其中miRNA 的含量，并分析其与肾纤维化的关系。结果：超速离心获得的尿液沉淀物呈现exosomes 的形态学特
征。miR-21、miR-29b、miR-29c、miR-30e、miR-192、miR-200a、miR-200c 和miR-429 可在所有患者的尿液exosomes 中被检出，但含
量存在较大差异。与对照组相比，纤维化组患者尿液exosomes 中，miR-21、miR-29b、miR-30e 和miR-200c 的含量显著增高，
miR-29c 的含量显著下降，而miR-192、miR-200a和miR-429 的含量无显著变化。尿液exosomes 中miRNA含量与纤维化肾组织
中miRNA表达量的变化并不完全一致。结论：尿液exosomes中miR-29c 和miR-21 的含量在肾纤维化的病变中发生显著改变，
可能成为早期诊断肾纤维化的生物标志物。 |
| 英文摘要: |
| Objective:Renal fibrosis is an inevitable outcome of chronic kidney disease (CKD). However, early diagnose of renal
fibrosis was difficult. It was reported in recent years that various microRNAs (miRNAs) regulate the progression of tubulointerstitial fibrosis.
In this study, whether changes of miRNA levels in urine exosomes were associated with renal fibrosis were evaluated.Methods:Morning urine specimens were collected frompatients suffered fromprimary glomerular diseases. Patients whose renal biopsy showed no
and mild tubulointerstitial fibrosis were enrolled in control and fibrosis group, respectively. Urine exosomes were isolated by ultracentrifugation
and observed by transmission electron microscopy. Quantifications of urinary miRNAs were performed using stem-loop
qRT-PCR followed by TaqMan PCR.Results:Isolated exosomes from urine samples appeared as clusters of vesicles of 30-100nm in diameter
under electron microscope, surrounded by a double-layer membrane. Most of previously reported fibrosis-associated miRNAs, including
miR-21, miR-29s(miR-29b and miR-29c), miR-30e, miR-192, miR-200s (miR-200a, miR-200c and miR-429) were detectable in
urine exosomes. However, their contents varied significantly. MiR-21, miR-29b, miR-30e and miR-200c were upregulated in urine exosomes,
while miR-29c was downregulated. Furthermore, miRNA levels in exosomes were not associated with their expression levels in
fibrotic kidney.Conclusion:Levels of miR-29c and miR-30e in urine exosomes changed significantly in the development of renal fibrosis,
which might indicate the renal tubulointerstitial fibrosis. |
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