MicroRNA-184 is a downstream effector of albuminuria driving renal fibrosis in rats with diabetic nephropathy.
MicroRNA-184 is a downstream effector of albuminuria driving renal fibrosis in rats with diabetic nephropathy.
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DOI：
10.1007/s00125-017-4248-9
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发表时间：
2017-06
期刊：
影响因子：
8.2
通讯作者：
Zoja C
中科院分区：
文献类型：
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作者：
Zanchi C;Macconi D;Trionfini P;Tomasoni S;Rottoli D;Locatelli M;Rudnicki M;Vandesompele J;Mestdagh P;Remuzzi G;Benigni A;Zoja C
肾纤维化是糖尿病肾病的常见并发症，也是导致终末期肾病的主要原因。尽管糖尿病肾病中肾纤维化与 microRNA (miRNA) 失调之间存在联系，但所涉及的特定 miRNA 的鉴定仍然不完整。本研究的目的是研究糖尿病肾脏中的 miRNA 谱，并确定与肾纤维化有关的潜在下游靶点。对患有明显肾病的 8 个月大 Zucker 糖尿病脂肪 (ZDF) 大鼠的肾脏中的 miRNA 表达谱进行了研究。通过原位杂交确定上调最多的 miRNA 的定位。通过计算机分析鉴定了候选 miRNA 靶标，并记录了其在糖尿病肾脏中与纤维化标志物相关的表达。培养的肾小管细胞用于评估哪种促纤维化刺激物可触发过度表达的 miRNA，并研究潜在的表观遗传机制。在 ZDF 大鼠中，与瘦大鼠相比，miR-184 显示出最强的差异上调（18 倍）。 miR-184 的肾小管定位与脂质磷酸磷酸酶 3 (LPP3) 表达减少和胶原蛋白积累相关。用 miR-184 模拟物转染 NRK-52E 细胞可减少 LPP3，促进促纤维化表型。白蛋白是 miR-184 表达的主要触发因素。抗 miR-184 抵消了白蛋白诱导的 LPP3 下调和纤溶酶原激活剂抑制剂 1 的过度表达。在 ZDF 大鼠中，ACE 抑制剂治疗限制了蛋白尿并减少了 miR-184，同时保留了肾小管 LPP3 并改善了肾小管间质纤维化。白蛋白诱导的肾小管细胞中的 miR-184 表达通过 DNA 去甲基化和组蛋白赖氨酸乙酰化进行表观遗传调节，并伴随着 NF-κB p65 亚基与 miR-184 启动子的结合。这些结果表明，miR-184可能通过LPP3作为蛋白尿的下游效应子，促进肾小管间质纤维化，并为研究靶向miR-184与降低蛋白尿药物联合是否可能是在糖尿病肾病中实现完全抗纤维化作用的新策略提供了理论基础。本文的在线版本 (doi:10.1007/s00125-017-4248-9) 包含经过同行评审但未经编辑的补充材料，可供授权用户使用。
Renal fibrosis is a common complication of diabetic nephropathy and is a major cause of end-stage renal disease. Despite the suggested link between renal fibrosis and microRNA (miRNA) dysregulation in diabetic nephropathy, the identification of the specific miRNAs involved is still incomplete. The aim of this study was to investigate miRNA profiles in the diabetic kidney and to identify potential downstream targets implicated in renal fibrosis. miRNA expression profiling was investigated in the kidneys of 8-month-old Zucker diabetic fatty (ZDF) rats during overt nephropathy. Localisation of the most upregulated miRNA was established by in situ hybridisation. The candidate miRNA target was identified by in silico analysis and its expression documented in the diabetic kidney associated with fibrotic markers. Cultured tubule cells served to assess which of the profibrogenic stimuli acted as a trigger for the overexpressed miRNA, and to investigate underlying epigenetic mechanisms. In ZDF rats, miR-184 showed the strongest differential upregulation compared with lean rats (18-fold). Tubular localisation of miR-184 was associated with reduced expression of lipid phosphate phosphatase 3 (LPP3) and collagen accumulation. Transfection of NRK-52E cells with miR-184 mimic reduced LPP3, promoting a profibrotic phenotype. Albumin was a major trigger of miR-184 expression. Anti-miR-184 counteracted albumin-induced LPP3 downregulation and overexpression of plasminogen activator inhibitor-1. In ZDF rats, ACE-inhibitor treatment limited albuminuria and reduced miR-184, with tubular LPP3 preservation and tubulointerstitial fibrosis amelioration. Albumin-induced miR-184 expression in tubule cells was epigenetically regulated through DNA demethylation and histone lysine acetylation and was accompanied by binding of NF-κB p65 subunit to miR-184 promoter. These results suggest that miR-184 may act as a downstream effector of albuminuria through LPP3 to promote tubulointerstitial fibrosis, and offer the rationale to investigate whether targeting miR-184 in association with albuminuria-lowering drugs may be a new strategy to achieve fully anti-fibrotic effects in diabetic nephropathy. The online version of this article (doi:10.1007/s00125-017-4248-9) contains peer-reviewed but unedited supplementary material, which is available to authorised users.