Silenced miR-21 inhibits renal interstitial fibrosis via targeting ERK1/2 signaling pathway in mice.
Silenced miR-21 inhibits renal interstitial fibrosis via targeting ERK1/2 signaling pathway in mice.
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DOI：
10.26355/eurrev_201908_18637
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发表时间：
2019-08
影响因子：
3.3
通讯作者：
Chuanyan Tang;Shihua Luo;X. Lin;J. Wang;Y. Liu
中科院分区：
文献类型：
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作者：
Chuanyan Tang;Shihua Luo;X. Lin;J. Wang;Y. Liu
目的研究微小核糖核酸（miR-21）对肾间质纤维化（RIF）模型小鼠的影响，通过研究miR-21对细胞外信号调节激酶（ERK）1/2信号通路相关蛋白及其下游蛋白表达的影响，初步阐明miR-21在RIF发生发展中的作用机制。材料与方法建立小鼠左侧输尿管梗阻模型。将实验小鼠分为假手术组和UUO组，正常饲养3周。然后处死，抽取血液测定肾功能相关指标。切除左肾，留取相应标本，观察肾脏外观及肾小管、肾小管形态。检测肾组织上皮钙粘蛋白（E-cadherin）、α-平滑肌肌动蛋白（α-SMA）、磷酸化ERK 1/2（p-ERK 1/2）、转化生长因子-β1（TGF-β1）和结缔组织生长因子（CTGF）蛋白的相对表达水平。人肾近端小管上皮细胞株HK-2经高糖（HG）联合沉默miR-21或ERK 1/2抑制剂PD 98059处理后，检测TGF-β1和β-SMA蛋白的相对表达水平。结果UUO组血尿素氮（BUN）、血清肌酐（SCr）、尿酸（UA）含量明显高于Sham组，肾间质单核细胞和淋巴细胞浸润，肾小管足细胞损伤、表型转化和萎缩，间质成纤维细胞活化增殖，细胞外基质（ECM）过度沉积。肾组织中E-cadherin的表达水平降低，而β-SMA、TGF-β1、CTGF和p-ERK 1/2蛋白的相对表达水平明显升高。HG与沉默miR-21或ERK 1/2抑制剂PD 98059联合处理后，人肾近端肾小管上皮细胞系HK-2中β-SMA和TGF-β1蛋白的相对表达水平降低。结论miR-21可能与RIF的发生发展有关。沉默的miR-21可能通过ERK 1/2信号通路抑制RIF。
OBJECTIVE To study the influence of micro ribonucleic acid (miR)-21 on the renal interstitial fibrosis (RIF) model mice, and to preliminarily elucidate the mechanism of action of miR-21 in the development of RIF by studying the influences of miR-21 on the expressions of the proteins related to the extracellular signal-regulated kinase (ERK) 1/2 signaling pathway and its downstream proteins. MATERIALS AND METHODS The mouse model of the left unilateral ureteral obstruction (UUO) was established. The experimental mice were divided into the Sham group and UUO group and were normally fed for 3 weeks. Then, they were executed, and their blood was extracted to determine the renal function-related indicators. The left kidney was excised, and the corresponding specimens were reserved for observing the appearance of the kidney and the morphology of the renal tubules and interstitium. The relative expression levels of epithelial (E-)cadherin, α-smooth muscle actin (α-SMA), and ERK1/2 phosphorylated ERK1/2 (p-ERK1/2), the transforming growth factor-β1 (TGF-β1) and the connective tissue growth factor (CTGF) proteins in renal tissues were determined. After the human renal proximal tubular epithelial cell line, the human kidney-2 (HK-2) was treated with high glucose (HG) combined with silenced miR-21 or the ERK1/2 inhibitor PD98059, the relative expression levels of ɑ-SMA and TGF-β1 protein were measured. RESULTS UUO group had significantly higher content of blood urea nitrogen (BUN), serum creatinine (SCr), and uric acid (UA) than the Sham group, and exhibited the infiltration of renal interstitial monocytes and lymphocytes, renal tubular podocyte damage, phenotypic transformation and atrophy, the activation and proliferation of interstitial fibroblasts, and excessive deposition of extracellular matrix (ECM). Moreover, the expression level of E-cadherin in the renal tissues was decreased, but the relative expression levels of ɑ-SMA, and TGF-β1, CTGF, and p-ERK1/2 proteins were evidently elevated. Lower relative expression levels of ɑ-SMA and TGF-β1 protein were detected in the human renal proximal tubular epithelial cell line HK-2 after the combined treatment with HG and silenced miR-21 or the ERK1/2 inhibitor PD98059. CONCLUSIONS MiR-21 may be related to the occurrence and development of RIF. Silenced miR-21 probably suppresses RIF via the ERK1/2 signaling pathway.