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.
引文
[1]Asrih M,Steffens S.Emerging role of epigenetics and miRNA in diabetic cardiomyopathy[J].Cardiovasc Pathol,2013,22(2):117-125.
[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.