微RNA-21通过激活沉默信息调控因子1信号通路缓解多柔比星心肌毒性
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摘要
目的探讨微RNA-21(miR-21)能否减轻多柔比星(DOX)心肌毒性,并阐明沉默信息调控因子1(SIRT1)信号通路是否介导其作用。方法用DOX(1μmol/L)处理大鼠原代心肌细胞构建DOX心肌毒性模型。将心肌细胞分为8组:对照组、miR-21组、miR-21抑制剂组、DOX组、miR-21+DOX组、miR-21抑制剂+DOX组、Sirtinol+mi R-21+DOX组、Sirtinol+DOX组,mi R-21 mimics、mi R-21抑制剂和Sirtinol(SIRT1抑制剂)分别于DOX处理前24 h加入细胞培养液中。DOX处理24 h后检测心肌细胞的细胞活力、凋亡率、凋亡相关蛋白和SIRT1信号通路表达情况。结果与对照组相比,DOX处理24 h后心肌细胞活力降低,Bcl-2和SIRT1表达量降低,而Bax和cleaved Caspase-3表达量增加,细胞凋亡率增高,差异均有统计学意义(P<0.05)。与DOX组相比,miR-21可明显提高心肌细胞活力,上调Bcl-2和SIRT1表达,下调Bax和cleaved Caspase-3表达,降低细胞凋亡率,差异均有统计学意义(P<0.05)。抑制SIRT1信号通路可削弱miR-21对心肌细胞的保护作用(P<0.05)。结论 mi R-21可通过激活SIRT1信号通路抑制心肌细胞凋亡,提高细胞活力,缓解DOX心肌毒性。
Objective To explore whether microR-21(miR-21) can alleviate doxorubicin(DOX)-induced cardiotoxicity and whether silent information regulator 1(SIRT1) signaling pathway mediates the roles. Methods Neonatal rat cardiac myocytes were treated with DOX(1 μmol/L) to induce DOX myocardial toxicity model. The cardiomyocytes were randomized into 8 groups: control group, miR-21 group, miR-21 inhibitor group, DOX group, miR-21+DOX group, miR-21 inhibitor+DOX group, Sirtinol+mi R-21+DOX group and Sirtinol+DOX group. The miR-21 mimics, miR-21 inhibitors and Sirtinol(SIRT1 inhibitor) were given at 24 h before DOX treatment. After treatment with DOX for 24 h, the cell viability, apoptosis rate, and the expression levels of apoptosis-related proteins and SIRT1 signaling pathway were detected. Results Compared with the control group, the cell viability, and the expression levels of Bcl-2 and SIRT1 were significantly decreased in the cardiomyocytes after treatment with DOX for 24 h, while the expression levels of Bax and cleaved Caspase-3, and apoptotic rate were significantly increased(P<0.05). Compared with the DOX group, miR-21 significantly increased cell viability and the expression levels of Bcl-2 and SIRT1, and significantly decreased the expression levels of Bax and cleaved Caspase-3 and apoptotic rate(P<0.05). Inhibiting SIRT1 signaling pathway could significantly weaken the protective effect of miR-21 on cardiomyocytes(P<0.05). Conclusion miR-21 can inhibit cardiomyocyte apoptosis, increase cell viability and alleviate DOX-induced cardiotoxicity by activating SIRT1 signaling pathway.
Objective To explore whether microR-21(miR-21) can alleviate doxorubicin(DOX)-induced cardiotoxicity and whether silent information regulator 1(SIRT1) signaling pathway mediates the roles. Methods Neonatal rat cardiac myocytes were treated with DOX(1 μmol/L) to induce DOX myocardial toxicity model. The cardiomyocytes were randomized into 8 groups: control group, miR-21 group, miR-21 inhibitor group, DOX group, miR-21+DOX group, miR-21 inhibitor+DOX group, Sirtinol+mi R-21+DOX group and Sirtinol+DOX group. The miR-21 mimics, miR-21 inhibitors and Sirtinol(SIRT1 inhibitor) were given at 24 h before DOX treatment. After treatment with DOX for 24 h, the cell viability, apoptosis rate, and the expression levels of apoptosis-related proteins and SIRT1 signaling pathway were detected. Results Compared with the control group, the cell viability, and the expression levels of Bcl-2 and SIRT1 were significantly decreased in the cardiomyocytes after treatment with DOX for 24 h, while the expression levels of Bax and cleaved Caspase-3, and apoptotic rate were significantly increased(P<0.05). Compared with the DOX group, miR-21 significantly increased cell viability and the expression levels of Bcl-2 and SIRT1, and significantly decreased the expression levels of Bax and cleaved Caspase-3 and apoptotic rate(P<0.05). Inhibiting SIRT1 signaling pathway could significantly weaken the protective effect of miR-21 on cardiomyocytes(P<0.05). Conclusion miR-21 can inhibit cardiomyocyte apoptosis, increase cell viability and alleviate DOX-induced cardiotoxicity by activating SIRT1 signaling pathway.
引文
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[2]TEMPFER C B,GIGER-PABST U,SEEBACHER V,PETERSEN M,DOGAN A,REZNICZEK G A.A phaseⅠ,single-arm,open-label,dose escalation study of intraperitoneal cisplatin and doxorubicin in patients with recurrent ovarian cancer and peritoneal carcinomatosis[J].Gynecol Oncol,2018,150:23-30.
[3]ZHAI Q,CHEN Y,XU J,HUANG Y,SUN J,LIU Y,et al.Lymphoma immunochemotherapy:targeted delivery of doxorubicin via a dual functional nanocarrier[J].Mol Pharm,2017,14:3888-3895.
[4]MINOTTI G,MENNA P,SALVATORELLI E,CAIRAG,GIANNI L.Anthracyclines:molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity[J].Pharmacol Rev,2004,56:185-229.
[5]ZHANG YW,SHI J,LI Y J,WEI L.Cardiomyocyte death in doxorubicin-induced cardiotoxicity[J].Arch Immunol Ther Exp(Warsz),2009,57:435-445.
[6]LI J,XU J,CHENG Y,WANG F,SONG Y,XIAO J.Circulating microRNAs as mirrors of acute coronary syndromes:miRacle or quagMire?[J].J Cell Mol Med,2013,17:1363-1370.
[7]LIU X,XIAO J,ZHU H,WEI X,PLATT C,DAMILANO F,et al.miR-222 is necessary for exerciseinduced cardiac growth and protects against pathological cardiac remodeling[J].Cell Metab,2015,21:584-595.
[8]CHEN J,HUANG Z P,SEOK H Y,DING J,KATAOKAM,ZHANG Z,et al.miR-17-92 cluster is required for and sufficient to induce cardiomyocyte proliferation in postnatal and adult hearts[J].Circ Res,2013,112:1557-1566.
[9]MELMAN Y F,SHAH R,DANIELSON K,XIAOJ,SIMONSON B,BARTH A,et al.Circulating microRNA-30d is associated with response to cardiac resynchronization therapy in heart failure and regulates cardiomyocyte apoptosis:a translational pilot study[J].Circulation,2015,131:2202-2216.
[10]WANG N,YANG J,ZHANG H,LU X,WANG J,CAO Y,et al.MicroRNA-9a-5p alleviates ischemia injury after focal cerebral ischemia of the rat by targeting ATG5-mediated autophagy[J].Cell Physiol Biochem,2018,45:78-87.
[11]DONG S,CHENG Y,YANG J,LI J,LIU X,WANG X,et al.MicroRNA expression signature and the role of microRNA-21 in the early phase of acute myocardial infarction[J].J Biol Chem,2009,284:29514-29525.
[12]VICZENCZOVA C,SZEIFFOVA BACOVA B,EGANBENOVA T,KURA B,YIN C,WEISMANN P,et al.Myocardial connexin-43 and PKC signalling are involved in adaptation of the heart to irradiation-induced injury:implication of miR-1 and miR-21[J].Gen Physiol Biophys,2016,35:215-222.
[13]YANG Q,YANG K,LI A.microRNA-21 protects against ischemia-reperfusion and hypoxia-reperfusion-induced cardiocyte apoptosis via the phosphatase and tensin homolog/Akt-dependent mechanism[J].Mol Med Rep,2014,9:2213-2220.
[14]TONG Z,JIANG B,WU Y,LIU Y,LI Y,GAO M,et al.MiR-21 protected cardiomyocytes against doxorubicininduced apoptosis by targeting BTG2[J].Int J Mol Sci,2015,16:14511-14525.
[15]RUAN Y,DONG C,PATEL J,DUAN C,WANG X,WU X,et al.SIRT1 suppresses doxorubicin-induced cardiotoxicity by regulating the oxidative stress and p38MAPK pathways[J].Cell Physiol Biochem,2015,35:1116-1124.
[16]LIN Q,GENG Y,ZHAO M,LIN S,ZHU Q,TIAN Z.MiR-21 regulates TNF-α-induced CD40 expression via the SIRT1-NF-κB pathway in renal inner medullary collecting duct cells[J].Cell Physiol Biochem,2017,41:124-136.
[17]ZHAI M,LIU Z,ZHANG B,JING L,LI B,LI K,et al.Melatonin protects against the pathological cardiac hypertrophy induced by transverse aortic constriction through activating PGC-1β:in vivo and in vitro studies[J/OL].J Pineal Res,2017,63:e12433.doi:10.1111/jpi.12433.
[18]ZHU S G,KUKREJA R C,DAS A,CHEN Q,LESNEFSKY E J,XI J.Dietary nitrate supplementation protects against doxorubicin-induced cardiomyopathy by improving mitochondrial function[J].J Am Coll Cardiol,2011,57:2181-2189.
[19]ZHAI M,LI B,DUAN W,JING L,ZHANG B,ZHANGM,et al.Melatonin ameliorates myocardial ischemia reperfusion injury through SIRT3-dependent regulation of oxidative stress and apoptosis[J/OL].J Pineal Res,2017,63:e12419.doi:10.1111/jpi.12419.
[20]DA COSTA MARTINS P A,LEPTIDIS S,SALIC K,DE WINDT L J.MicroRNA regulation in cardiovascular disease[J].Curr Drug Targets,2010,11:900-906.
[21]SCHULTE C,KARAKAS M,ZELLER T.microRNAs in cardiovascular disease―clinical application[J].Clin Chem Lab Med,2017,55:687-704.
[22]VIJAYARATHNA S,OON C E,JOTHY S L,CHEN Y,KANWAR J R,SASIDHARAN S.MicroRNA pathways:an emerging role in identification of therapeutic strategies[J].Curr Gene Ther,2014,14:112-120.
[23]CHENG Y,ZHANG C.MicroRNA-21 in cardiovascular disease[J].J Cardiovasc Transl Res,2010,3:251-255.
[24]GUPTA S K,ITAGAKI R,ZHENG X,BATKAI S,THUM S,AHMAD F,et al.miR-21 promotes fibrosis in an acute cardiac allograft transplantation model[J].Cardiovasc Res,2016,110:215-226.
[25]DAI B,LI H,FAN J,ZHAO Y,YIN Z,MIE X,et al.MiR-21 protected against diabetic cardiomyopathy induced diastolic dysfunction by targeting gelsolin[J/OL].Cardiovasc Diabetol,2018,17:123.doi:10.1186/s12933-018-0767-z.
[26]肖静,潘宇,李晓红,杨翔宇,姜霖,冯娟,等.心脏干细胞来源的exosomes通过传递miR-21抗心肌细胞凋亡[J].中国病理生理杂志,2016,32:1533.
[27]D’ONOFRIO N,SERVILLO L,BALESTRIERI M L.SIRT1 and SIRT6 signaling pathways in cardiovascular disease protection[J].Antioxid Redox Signal,2018,28:711-732.
[28]CUI L,GUO J,ZHANG Q,YIN J,LI J,ZHOUW,et al.Erythropoietin activates SIRT1 to protect human cardiomyocytes against doxorubicin-induced mitochondrial dysfunction and toxicity[J].Toxicol Lett,2017,275:28-38.
[29]YUAN Y P,MA Z G,ZHANG X,XU S C,ZENG X F,YANG Z,et al.CTRP3 protected against doxorubicininduced cardiac dysfunction,inflammation and cell death via activation of Sirt1[J].J Mol Cell Cardiol,2018,114:38-47.
[30]OYAMA Y,BARTMAN C M,GILE J,ECKLE T.Circadian microRNAs in cardioprotection[J].Curr Pharm Des,2017,23:3723-3730.
[31]HUANG Y,YANG Y B,ZHANG X H,YU X L,WANGZ B,CHENG X C.MicroRNA-21 gene and cancer[J/OL].Med Oncol,2013,30:376.doi:10.1007/s12032-012-0376-8.
[32]BICA-POP C,COJOCNEANU-PETRIC R,MAGDOL,RADULY L,GULEI D,BERINADAN-NEAGEOL.Overview upon miR-21 in lung cancer:focus on NSCLC[J].Cell Mol Life Sci,2018,75:3539-3551.
[33]初旭,刘永兴,党新文,周永红,赵咪.microRNA-21对人舌鳞癌细胞的增殖和凋亡的影响[J].现代生物医学进展,2018,18:1940-1943.
[34]ZHANG S,HAN L,WEI J,SHI Z,PU P,ZHANGJ,et al.Combination treatment with doxorubicin and microRNA-21 inhibitor synergistically augments anticancer activity through upregulation of tumor suppressing genes[J].Int J Oncol,2015,46:1589-1600.