Silencing of microRNA-132 reduces renal fibrosis by selectively inhibiting myofibroblast proliferation
Silencing of microRNA-132 reduces renal fibrosis by selectively inhibiting myofibroblast proliferation
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DOI：
10.1016/j.kint.2016.01.029
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发表时间：
2016-06-01
影响因子：
19.6
通讯作者：
van Zonneveld, Anton Jan
中科院分区：
文献类型：
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作者：
Bijkerk, Roel;de Bruin, Ruben G.;van Zonneveld, Anton Jan
慢性肾脏疾病与进行性肾脏纤维化有关，血管周围细胞产生大多数α-平滑肌肌动蛋白(α-SMA)阳性的肌成纤维细胞。在这里，我们试图确定可以作为减少肌成纤维细胞形成的靶点的周细胞性miRNAs。通过单侧输尿管梗阻诱导FoxD1-GC；Z/Red小鼠肾脏纤维化，然后对dsRed阳性的FoxD1-派生细胞进行FACS分选和miRNA图谱分析。在周细胞到肌成纤维细胞的形成过程中，miR-132选择性地增加了21倍，而miR-132在总肾脏裂解物中只增加了2.5倍(无论是在梗阻还是在缺血-再灌注损伤中)。MIR-132在梗阻过程中的沉默减少了胶原沉积(35%)和肾小管细胞凋亡。免疫组织化学、Western印迹和qRT-PCR证实间质α-SMA(+)细胞也有类似的减少。通路分析证实了miR-132在肌成纤维细胞增殖中的限速作用，这一点在体外得到证实。事实上，用antagomir-132治疗的小鼠表现出增殖的Ki67(+)间质肌成纤维细胞的数量减少。有趣的是，这对间质有选择性，并不损害肾小管上皮细胞的修复性增殖，表现为Ki67(+)上皮细胞的增加，以及肾脏裂解物中磷酸化RB1、Cyclin-A的增加和RASA1、p21水平的降低。其他途径和基因表达分析表明，miR-132协调调节参与转化生长因子-β信号转导(Smad2/SMAD3)、STAT3/ERK通路和细胞增殖(Foxo3/p300)的基因。因此，沉默miR-132通过选择性地抑制肌成纤维细胞的增殖来对抗肾脏纤维化的进展，并可能成为一种新的抗纤维化治疗方法。
Chronic kidney disease is associated with progressive renal fibrosis, where perivascular cells give rise to the majority of alpha-smooth muscle actin (alpha-SMA) positive myofibroblasts. Here we sought to identify pericytic miRNAs that could serve as a target to decrease myofibroblast formation. Kidney fibrosis was induced in FoxD1-GC; Z/Red-mice by unilateral ureteral obstruction followed by FACS sorting of dsRed-positive FoxD1-derivative cells and miRNA profiling. MiR-132 selectively increased 21-fold during pericyte-to-myofibroblast formation, whereas miR-132 was only 2.5-fold up in total kidney lysates (both in obstructive and ischemia-reperfusion injury). MiR-132 silencing during obstruction decreased collagen deposition (35%) and tubular apoptosis. Immunohistochemistry, Western blot, and qRT-PCR confirmed a similar decrease in interstitial alpha-SMA(+) cells. Pathway analysis identified a rate-limiting role for miR-132 in myofibroblast proliferation that was confirmed in vitro. Indeed, antagomir-132-treated mice displayed a reduction in the number of proliferating Ki67(+) interstitial myofibroblasts. Interestingly, this was selective for the interstitial compartment and did not impair the reparative proliferation of tubular epithelial cells, as evidenced by an increase in Ki67(+) epithelial cells, as well as increased phospho-RB1, Cyclin-A and decreased RASA1, p21 levels in kidney lysates. Additional pathway and gene expression analyses suggest miR-132 coordinately regulates genes involved in TGF-beta signaling (Smad2/Smad3), STAT3/ERK pathways, and cell proliferation (Foxo3/p300). Thus, silencing miR-132 counteracts the progression of renal fibrosis by selectively decreasing myofibroblast proliferation and could potentially serve as a novel antifibrotic therapy.