高糖对H9C2心肌细胞miR?26b表达的影响
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摘要
目的糖尿病心肌病(DCM)是糖尿病的并发症之一,miRNA的改变与DCM的发生发展密切相关,文中观察了C57BL/6J小鼠组织中miR?26b的表达特征,ob/ob小鼠心脏、白色脂肪组织及肝内miR?26b水平的变化以及高糖对H9C2心肌细胞中miR?26b表达的影响。方法以C57BL/6J小鼠及ob/ob小鼠为研究对象,采用RT?PCR法检测C57BL/6J小鼠以及ob/ob小鼠心脏、脂肪、肝等组织中miR?26b水平;另以H9C2心肌细胞为工具,分为5.5、15、25、35 mmol/L葡萄糖(分别定义为5.5、15、25、35 mmol/L葡萄糖组)干预0、24、48、72、96、120 h,采用CCK?8法检测细胞增殖变化;另采用RT?PCR法检测4组细胞中miR?26b水平。结果在C57BL/6J小鼠心脏组织中miR?26b表达水平最高;与野生型C57BL/6J小鼠相比,ob/ob小鼠心脏及白色脂肪组织中miR?26b水平明显降低(P<0.05);与5.5 mmol/L葡萄糖组比较,15、25及35 mmol/L 3组细胞24、48、72、96、120h增殖速度均显著增快(P<0.05);与15mmol/L葡萄糖组比较,25mmol/L葡萄糖组24 h增殖速度较快(0.72±0.01 vs 0.74±0.02,P<0.05),35mmol/L葡萄糖组48h增殖速度明显降低(0.94±0.01 vs 0.92±0.01,P<0.05),25、35mmol/L葡萄糖组96h增殖速度明显降低(P<0.05),35mmol/L葡萄糖组120h增殖速度稍有降低(1.19±0.05 vs 1.12±0.02,P<0.05);与25mmol/L葡萄糖组比较,35mmol/L葡萄糖组24、48h增殖速度均降低(P<0.05)。与5.5 mmol/L葡萄糖组比较,25、35 mmol/L葡萄糖组细胞中miR?26b水平显著下调(P <0.05)。25mmol/L葡萄糖干预后,与0h miR?26b表达水平比较,96、120hmiR?26b表达水平明显降低(P<0.05)。结论高浓度葡萄糖可诱导心肌细胞中miR?26b表达水平降低,miR?26b可能参与介导DCM的发病过程。
Objective Diabetic cardiomyopathy(DCM)is one of the complications of diabetes,which is closely related to thechange of miRNA. In this study,we observed the characteristic expression of miR-26 b in the tissues of the C57 BL/6 J mouse and in theheart,adipose tissue and liver of the ob/ob mouse,and investigated the effect of high glucose(Glu)on the expression of miR-26 b in H9 C2 cardiomyocytes.Methods Using RT-PCR,we measured the levels of miR-26 b in the heart,adipose tissue,liver and other tissues ofC57 BL/6 J and ob/ob mice. H9 C2 cardiomyocytes were treated with Glu at 5.5,15,25 and 35 mmol/L for 0,24,48,72,96 and 120 hours,followed by detection of the proliferation of cardiomyocytes by CCK-8 and determination of thelevels miR-26 b.Results The expression of miR-26 b was the highest in the heart of the C57 BL/6 J mice,significantly higher than in the cardiac and white adipose tissues of the ob/ob mice(P < 0.05). The proliferation of cardiomyocytes was markedly increased in the 15,25 and 35 mmol/L Glu groups at 24,48,72,96 and 120 hours as compared with that in the 5.5 mmol/L Glu group(P < 0.05),higher in the 25 than in the 15 mmol/L Glu group at 24 hours(0.74±0.02 vs 0.72±0.01,P<0.05),but lower in the 35 than in the 15 mmol/L Glu group at 48 hours(0.92±0.01 vs 0.94±0.01,P<0.05),in the 25 and 35 mmol/LGlu groups at 96 hours(P < 0.05),in the 35 mmol/L Glu group at 120 hours(1.12±0.02 vs 1.19±0.05,P<0.05),in the 35 than in the 25 mmol/L Glu group at 24 and 48 hours(P<0.05). The expression of miR-26 b in the H9 C2 cardiomyocytes was significantly down-regulated inthe 25 and 35 mmol/L Glu groups in comparison with that in the 5.5 mmol/L Glu group(P<0.05),remarkably lower in the 25 mmol/L Glugroup at 96 and 120 hours than at 0 hour(P<0.05).Conclusion High glucose can down-regulate the expression of miR-26 b in H9 C2 car-diomyocytes,which suggests that miR-26 b may participate in the pathogenesis of DCM.
Objective Diabetic cardiomyopathy(DCM)is one of the complications of diabetes,which is closely related to thechange of miRNA. In this study,we observed the characteristic expression of miR-26 b in the tissues of the C57 BL/6 J mouse and in theheart,adipose tissue and liver of the ob/ob mouse,and investigated the effect of high glucose(Glu)on the expression of miR-26 b in H9 C2 cardiomyocytes.Methods Using RT-PCR,we measured the levels of miR-26 b in the heart,adipose tissue,liver and other tissues ofC57 BL/6 J and ob/ob mice. H9 C2 cardiomyocytes were treated with Glu at 5.5,15,25 and 35 mmol/L for 0,24,48,72,96 and 120 hours,followed by detection of the proliferation of cardiomyocytes by CCK-8 and determination of thelevels miR-26 b.Results The expression of miR-26 b was the highest in the heart of the C57 BL/6 J mice,significantly higher than in the cardiac and white adipose tissues of the ob/ob mice(P < 0.05). The proliferation of cardiomyocytes was markedly increased in the 15,25 and 35 mmol/L Glu groups at 24,48,72,96 and 120 hours as compared with that in the 5.5 mmol/L Glu group(P < 0.05),higher in the 25 than in the 15 mmol/L Glu group at 24 hours(0.74±0.02 vs 0.72±0.01,P<0.05),but lower in the 35 than in the 15 mmol/L Glu group at 48 hours(0.92±0.01 vs 0.94±0.01,P<0.05),in the 25 and 35 mmol/LGlu groups at 96 hours(P < 0.05),in the 35 mmol/L Glu group at 120 hours(1.12±0.02 vs 1.19±0.05,P<0.05),in the 35 than in the 25 mmol/L Glu group at 24 and 48 hours(P<0.05). The expression of miR-26 b in the H9 C2 cardiomyocytes was significantly down-regulated inthe 25 and 35 mmol/L Glu groups in comparison with that in the 5.5 mmol/L Glu group(P<0.05),remarkably lower in the 25 mmol/L Glugroup at 96 and 120 hours than at 0 hour(P<0.05).Conclusion High glucose can down-regulate the expression of miR-26 b in H9 C2 car-diomyocytes,which suggests that miR-26 b may participate in the pathogenesis of DCM.
引文
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[8]Raut SK,Singh GB,Rastogi B,et al.miR-30c and miR-181a synergistically modulate p53-p21 pathway in diabetes induced cardiac hypertrophy[J].Mol Cell Biochem,2016,417(1-2):191-203.
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[10]Jeyabal P,Thandavarayan RA,Joladarashi D,et al.MicroRNA-9 inhibits hyperglycemia-induced pyroptosis in human ventricular cardiomyocytes by targeting ELAVL1[J].Biochem Biophys Res Commun,2016,471(4):423-429.
[11]Xu G,Shi C,Ji C,et al.Expression of microRNA-26b,an obesity-related microRNA,is regulated by free fatty acids,glucose,dexamethasone and growth hormone in human adipocytes[J].Mol Med Rep,2014,10(1):223-228.
[12]Han M,Yang Z,Sayed D,et al.GATA4 expression is primarily regulated via a miR-26b-dependent post-transcriptional mechanism during cardiac hypertrophy[J].Cardiovasc Res,2012,93(4):645-654.
[13]Zhang ZH,Li J,Liu BR,et al.MicroRNA-26 was decreased in rat cardiac hypertrophy model and may be a promising therapeutic target[J].J Cardiovasc Pharm,2013,62(3):312-319.
[14]Rubi?P,Totoń?urańska J,Wi?niowska?mia?ek S,et al.Relations between circulating microRNAs(miR-21,miR-26,miR-29,miR-30 and miR-133a),extracellular matrix fibrosis and serum markers of fibrosis in dilated cardiomyopathy[J].Int JCardiol,2017,231:201-206.
[15]Xu G,Ji C,Song G,et al.MiR-26b modulates insulin sensitivity in adipocytes by interrupting the PTEN/PI3K/AKT pathway[J].Int J Obesity,2015,39(10):1523-1530.
[16]Xu G,Ji C,Song G,et al.Obesity-associated microRNA-26b regulates the proliferation of human preadipocytes via arrest of the G1/S transition[J].Mol Med Rep,2015,12(3):3648-3654.
[17]Song,G,Xu G,Ji C,et al.The role of microRNA-26b in human adipocyte differentiation and proliferation[J].Gene,2014,533(2):481-487.
[2]Wan A,Rodrigues B.Endothelial cell-cardiomyocyte crosstalk in diabetic cardiomyopathy[J].Cardiovasc Res,2016,111(3):172-183.
[3]唐香,江时森.糖尿病心肌病心肌线粒体膜分子异常的研究进展[J].医学研究生学报,2009,22(9):1005-1008.
[4]Guo R,Nair S.Role of microRNA in diabetic cardiomyopathy:From mechanism to intervention[J].Biochim Biophys Acta,2017,1863(8):2070-2077.
[5]胡蕴,毛晓明.miRNA在2型糖尿病中的研究进展[J].医学研究生学报,2011,24(9):1003-1006.
[6]Shenoy A,Blelloch RH.Regulation of microRNA function in somatic stem cell proliferation and differentiation[J].Nat Rev Mol Cell Biol,2014,15(9):565-576.
[7]康波,刘红明,洪江,等.硫化氢调节miRNA-455表达抑制内质网应激介导的心肌细胞凋亡[J].医学研究生学报,2014,27(12):1245-1249.
[8]Raut SK,Singh GB,Rastogi B,et al.miR-30c and miR-181a synergistically modulate p53-p21 pathway in diabetes induced cardiac hypertrophy[J].Mol Cell Biochem,2016,417(1-2):191-203.
[9]Zheng D,Ma J,Yu Y,et al.Silencing of miR-195 reduces diabetic cardiomyopathy in C57BL/6 mice[J].Diabetologia,2015,58(8):1949-1958.
[10]Jeyabal P,Thandavarayan RA,Joladarashi D,et al.MicroRNA-9 inhibits hyperglycemia-induced pyroptosis in human ventricular cardiomyocytes by targeting ELAVL1[J].Biochem Biophys Res Commun,2016,471(4):423-429.
[11]Xu G,Shi C,Ji C,et al.Expression of microRNA-26b,an obesity-related microRNA,is regulated by free fatty acids,glucose,dexamethasone and growth hormone in human adipocytes[J].Mol Med Rep,2014,10(1):223-228.
[12]Han M,Yang Z,Sayed D,et al.GATA4 expression is primarily regulated via a miR-26b-dependent post-transcriptional mechanism during cardiac hypertrophy[J].Cardiovasc Res,2012,93(4):645-654.
[13]Zhang ZH,Li J,Liu BR,et al.MicroRNA-26 was decreased in rat cardiac hypertrophy model and may be a promising therapeutic target[J].J Cardiovasc Pharm,2013,62(3):312-319.
[14]Rubi?P,Totoń?urańska J,Wi?niowska?mia?ek S,et al.Relations between circulating microRNAs(miR-21,miR-26,miR-29,miR-30 and miR-133a),extracellular matrix fibrosis and serum markers of fibrosis in dilated cardiomyopathy[J].Int JCardiol,2017,231:201-206.
[15]Xu G,Ji C,Song G,et al.MiR-26b modulates insulin sensitivity in adipocytes by interrupting the PTEN/PI3K/AKT pathway[J].Int J Obesity,2015,39(10):1523-1530.
[16]Xu G,Ji C,Song G,et al.Obesity-associated microRNA-26b regulates the proliferation of human preadipocytes via arrest of the G1/S transition[J].Mol Med Rep,2015,12(3):3648-3654.
[17]Song,G,Xu G,Ji C,et al.The role of microRNA-26b in human adipocyte differentiation and proliferation[J].Gene,2014,533(2):481-487.