LncRNA DCRF regulates cardiomyocyte autophagy by targeting miR-551b-5p in diabetic cardiomyopathy
LncRNA DCRF regulates cardiomyocyte autophagy by targeting miR-551b-5p in diabetic cardiomyopathy
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LncRNA DCRF 通过靶向 miR-551b-5p 调节糖尿病心肌病中的心肌细胞自噬
DOI：
10.7150/thno.31052
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发表时间：
2019-01-01
期刊：
影响因子：
12.4
通讯作者：
Zhou, Xiang
中科院分区：
文献类型：
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作者：
Feng, Yu;Xu, Weiting;Zhou, Xiang
背景：我们建立了糖尿病心肌病（DCM）大鼠模型，并报道了长链非编码RNA DCRF的显著上调。本研究旨在探讨DCRF在扩张型心肌病发生发展中的分子机制。方法：采用实时荧光定量PCR和RNA荧光原位杂交技术检测DCRF在心肌细胞中的表达。组织学和超声心动图分析用于评估DCRF敲低对糖尿病大鼠心脏结构和功能的影响。进行mRFP-GFP-LC 3荧光显微镜、透射电子显微镜和Western印迹以确定心肌细胞自噬。通过RNA免疫沉淀和荧光素酶报告基因分析来阐明DCRF/miR-551 b-5 p/PCDH 17通路在心肌细胞自噬中的调节作用。结果如下：我们的研究结果表明，DCRF敲低减少心肌细胞自噬，减轻心肌纤维化，改善糖尿病大鼠的心功能。高糖增加心肌细胞DCRF表达，诱导自噬。RNA免疫沉淀和荧光素酶报告基因分析表明，miR-551 b-5 p以AGO 2依赖的方式靶向DCRF，而PCDH 17是miR-551 b-5 p的直接靶点。发现DCRF的强制表达减弱了miR-551 b-5 p对PCDH 17的抑制作用。此外，DCRF敲低降低PCDH 17的表达，并抑制自噬在心肌细胞与高糖处理。结论：我们的研究表明，DCRF可以作为一种竞争性内源性RNA，通过吸收miR-551 b-5 p来增加PCDH 17的表达，从而有助于DCM中心肌细胞自噬的增加。
Background: We generated a rat model of diabetic cardiomyopathy (DCM) and reported significant upregulation of the long non-coding RNA DCRF. This study was designed to determine the molecular mechanisms of DCRF in the development of DCM. Methods: Real-time PCR and RNA fluorescent in situ hybridization were conducted to detect the expression pattern of DCRF in cardiomyocytes. Histological and echocardiographic analyses were used to assess the effect of DCRF knockdown on cardiac structure and function in diabetic rats. mRFP-GFP-LC3 fluorescence microscopy, transmission electron microscopy, and Western blotting were carried out to determine cardiomyocyte autophagy. RNA immunoprecipitation and luciferase reporter assays were performed to elucidate the regulatory role of DCRF/miR-551b-5p/PCDH17 pathway in cardiomyocyte autophagy. Results: Our findings showed that DCRF knockdown reduced cardiomyocyte autophagy, attenuated myocardial fibrosis, and improved cardiac function in diabetic rats. High glucose increased DCRF expression and induced autophagy in cardiomyocytes. RNA immunoprecipitation and luciferase reporter assays indicated that DCRF was targeted by miR-551b-5p in an AGO2-dependent manner and PCDH17 was the direct target of miR-551b-5p. Forced expression of DCRF was found to attenuate the inhibitory effect of miR-551b-5p on PCDH17. Furthermore, DCRF knockdown decreased PCDH17 expression and suppressed autophagy in cardiomyocytes treated with high glucose. Conclusion: Our study suggests that DCRF can act as a competing endogenous RNA to increase PCDH17 expression by sponging miR-551b-5p, thus contributing to increased cardiomyocyte autophagy in DCM.