基于miR-21/Smad7探讨芍药苷在抗血管紧张素Ⅱ诱导心肌肥大中的作用-东南大学学报医学版

基于miR-21/Smad7探讨芍药苷在抗血管紧张素Ⅱ诱导心肌肥大中的作用

单位：1. 河南省中医院心病科, 河南郑州 450000;
2. 河南中医药大学基础医学院, 河南郑州 450000;
3. 河南中医药大学研究生院, 河南郑州 450000

分类号：R-33;R541

出版年·卷·期（页码）：2018·37·第二期（191-197）

摘要：

目的：探讨芍药苷（Pae）在对抗血管紧张素Ⅱ（Ang-Ⅱ）诱导心肌肥大中的作用及其机制。方法：Ang-Ⅱ诱导建立心肌细胞肥大模型，分为正常对照组、Ang-Ⅱ组、Pae组及Ang-Ⅱ+Pae组，脂质体转染法将微小RNA-21模拟物（miR-21 mimic）、信号转导分子7（Smad7） siRNA及其对照转入模型细胞。图像分析系统测算心肌细胞相对表面积，实时PCR检测miR-21及心房利尿钠肽（ANP）、脑利尿钠肽（BNP） mRNA表达，双荧光素酶实验检测miR-21对Smad7的靶向作用。Western blotting观察Smad7、ANP及BNP蛋白表达。结果：Ang-Ⅱ诱导显著增加心肌细胞表面积，ANP、BNP mRNA及蛋白表达，上调miR-21，Pae能部分逆转Ang-Ⅱ诱导的miR-21表达增加（P<0.05），降低心肌细胞表面积，ANP、BNP mRNA及蛋白相对表达量（均P<0.05）。MiR-21靶向Smad7 3'-UTR，发挥对Smad7转录后翻译的抑制作用，Pae能降低miR-21对Smad7的抑制（P<0.05），Smad7敲除后削弱了Pae对心肌细胞作用的相对表面积以及对ANP、BNP mRNA及蛋白表达的抑制作用（均P<0.05）。结论：Pae能通过负性调节miR-21降低靶蛋白Smad7表达，发挥抵抗心肌细胞肥大的作用。

Objective:To identify the role of paeoniflorin(Pae) on attenuating Ang-Ⅱ induced cardiomyocyte hypertrophy. Methods:A model of cardiac hypertrophy induced by Ang-Ⅱ in vitro was developed. Cardiomyocytes were separated and divided into four groups:control group, Ang-Ⅱ group, Pae group and Ang-Ⅱ plus Pae group. Cells were transfected with miR-21 mimic, Smad7 siRNA or their negative control group. Myocardial cell image analysis system was used to measure the relative surface area of cardiomyocytes; Real-time PCR was used to assay the mRNA expression of miR-21, ANP and BNP. A dual luciferase reporter assay was used to detect the relationship between Smad7 activity and miR-21 expression.Western blotting was used to evaluate the protein level of Smad7, ANP and BNP. Results:Ang Ⅱ pretreatment significantly increased the myocardial cell surface area, as well as up-regulated the miR-21, ANP and BNP expression, while paeoniflorin can partly reverse these effects brought by Ang Ⅱ (all P<0.05). Results of dual luciferase reporter assay showed that miR-21 directly targeted the 3'-UTR of Smad7, resulting in the post-transcriptional translation inhibition of Smad7 (P<0.05). Pae can partly reduce the inhibition effect of miR-21 on Smad7 (P<0.05), Smad7 knockdown remarkably attenuated the inhibition effect on cell relative surface area, ANP and BNP expression induced by Pae (P<0.05). Conclusion:These results indicate that Pae may resistance to Ang-Ⅱ induced cardiac hypertrophy via miR-21/Smad7 pathway.

参考文献：

[1] BELL D,CAMPBELL M,WANG X,et al.Adrenomedullin gene delivery is cardio-protective in a model of chronic nitric oxide deficiency combining pressure overload, oxidative stress and cardiomyocyte hypertrophy[J].Cell Physiol Biochem, 2010,26(3):383-394.
[2] 于迎根,彭颖秀,史咏秋.慢性心衰患者心率减速力与室性心律失常的相关性研究[J].现代医学,2016,44(8):1074-1076.
[3] ZHAI J,GUO Y.Paeoniflorin attenuates cardiac dysfunction in endotoxemic mice via the inhibition of nuclear factor-kappaB[J].Biomed Pharmacother,2016,80:200-206.
[4] HAN F,ZHOU D,YIN X,et al.Paeoniflorin protects diabetic mice against myocardial ischemic injury via the transient receptor potential vanilloid 1/calcitonin gene-related peptide pathway[J].Cell Biosci,2016,6:37-40.
[5] YAN M,CHEN C,GONG W,et al.miR-21-3p regulates cardiac hypertrophic response by targeting histone deacetylase-8[J].Cardiovasc Res,2015,105(3):340-352.
[6] BANG C,BATKAI S,DANGWAL S,et al.Cardiac fibroblast-derived microRNA passenger strand-enriched exosomes mediate cardiomyocyte hypertrophy[J].J Clin Invest,2014,124(5):2136-2146.
[7] MOSTERD A,HOES A W.Clinical epidemiology of heart failure[J].Heart,2007, 93(9):1137-1146.
[8] 郑世存,李晓宇,欧阳兵,等.芍药苷药理作用研究新进展[J].中国药物警戒, 2012(2):100-103.
[9] QIAN G Q,DING J,ZHANG X,et al.Preconditioning with glycyrrhizic, ferulic, paeoniflorin, cinnamic prevents rat hearts from ischemia/reperfusion injury via endothelial nitric oxide pathway[J].Pharmacogn Mag,2015,11(42):292-296.
[10] ZHOU H,YANG H X,YUAN Y,et al.Paeoniflorin attenuates pressure overload-induced cardiac remodeling via inhibition of TGFbeta/Smads and NF-kappaB pathways[J].J Mol Histol,2013,44(3):357-367.
[11] OLIVEIRA-CARVALHO V,CARVALHO V O,SILVA M M,et al. MicroRNAs:a new paradigm in the treatment and diagnosis of heart failure?[J].Arq Bras Cardiol, 2012,98(4):362-369.
[12] THUM T,GROSS C,FIEDLER J,et al.MicroRNA-21 contributes to myocardial disease by stimulating MAP kinase signalling in fibroblasts[J].Nature,2008, 456(7224):980-984.
[13] SAYED D,RANE S,LYPOWY J,et al.MicroRNA-21 targets Sprouty2 and promotes cellular outgrowths[J].Mol Biol Cell,2008,19(8):3272-3282.
[14] BRONNUM H,ANDERSEN D C,SCHNEIDER M,et al.miR-21 promotes fibrogenic epithelial-to-mesenchymal transition of epicardial mesothelial cells involving. Programmed Cell Death 4 and Sprouty-1[J].PLoS One, 2013,8(2):e56280.
[15] JI Y,DOU Y N,ZHAO Q W,et al.Paeoniflorin suppresses TGF-beta mediated epithelial-mesenchymal transition in pulmonary fibrosis through a Smad-dependent pathway[J]. Acta Pharmacol Sin,2016,37(6):794-804.

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