title: MiRNA-30e mediated cardioprotection of ACE2 in rats with Doxorubicin-induced heart failure through inhibiting cardiomyocytes autophagy.
abstract: OBJECTIVE
  miRNAs are a class of small non-coding RNAs that has been proved to be involved in cardioprotection. The present study was to detect role of miR-30e in cardiac-protective action of ACE2 (angiotensin-converting enzyme 2).
  
  
  METHODS
  Sprague Dawley rats were divided into 3 groups and received treatment for a total of 6weeks: group1, normal rats; group2, Doxorubicin-induced heart cardiomyopathy (DHC) rats; and group3, rhACE2 (recombinant human ACE2) treated DHC rats. Doxorubicin was discontinuously administered via intraperitoneal injection. Primary cardiomyocytes and H9C2 cell line were used for in vitro experiments. MiR-30a, miR-30c and miR-30e expression were determined using qRT-PCR. Expression of autophagy associated gene expression including Beclin-1 and LC3 II/I were determined using western blot. Cell apoptosis was evaluated using TUNEL assay.
  
  
  RESULTS
  Administration of ACE2 suppressed harmful action of Doxorubicin and caused a significant improvement of left ventricular contractility function, upregulation in miR-30 (a, c and e) expression, and inhibition in Beclin-1 expression and LC3-II/I ratio. This was supported by results of Ad-ACE2-incubated primary cardiomyocytes. By manipulating miR-30e expression in H9C2 cells, we observed that miR-30e regulated Beclin-1 expression via inhibiting its 3'UTR activity. MiR-30e mimic treatment resulted in downregulation of Beclin-1 and protected primary cardiomyocytes against apoptosis. Moreover, silencing miR-30e induced cardiomyocytes apoptosis was abrogated by ACE2 overexpresssion. This was further confirmed by in vivo DHC rat experiments that showed that co-injected ACE2 and miR-30 inhibitor reduced cardiac function.
  
  
  CONCLUSIONS
  In summary, administration of ACE2 attenuates Doxorubicin-induced cardiac dysfunction via preservation of cardiomyocytes autophagy in a miR-30e/beclin-1 signal pathway.
authors.affiliation: Department of Cardiology, the First People's Hospital of Hangzhou, Hangzhou 310006, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Lai
authors.affiliation: Department of Cardiology, the First People's Hospital of Hangzhou, Hangzhou 310006, China. Electronic address: c_junzhu@163.com.
authors.forename: Junzhu
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Cardiology, the First People's Hospital of Hangzhou, Hangzhou 310006, China.
authors.forename: Ningfu
authors.initials: N
authors.lastname: Wang
authors.affiliation: Department of Cardiology, the First People's Hospital of Hangzhou, Hangzhou 310006, China.
authors.forename: Gangjie
authors.initials: G
authors.lastname: Zhu
authors.affiliation: Department of Cardiology, the First People's Hospital of Hangzhou, Hangzhou 310006, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Duan
authors.affiliation: Department of Cardiology, the First People's Hospital of Hangzhou, Hangzhou 310006, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Ling
mesh_terms: D000085962:Angiotensin-Converting Enzyme 2; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000009:adverse effects*; D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy / Q000187:drug effects; D000071186:Beclin-1 / Q000235:genetics; D009202:Cardiomyopathies / Q000139:chemically induced* / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D002316:Cardiotonic Agents / Q000627:therapeutic use*; D002460:Cell Line; D002478:Cells, Cultured; D004317:Doxorubicin / Q000009:adverse effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D007703:Peptidyl-Dipeptidase A / Q000627:therapeutic use*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000627:therapeutic use
chemical_list: D000903:Antibiotics, Antineoplastic; D000071186:Beclin-1; D002316:Cardiotonic Agents; C556157:MIRN30 microRNA, rat; D035683:MicroRNAs; D011994:Recombinant Proteins; D004317:Doxorubicin; D007703:Peptidyl-Dipeptidase A; C000705307:ACE2 protein, human; C000705329:Ace2 protein, rat; D000085962:Angiotensin-Converting Enzyme 2
keywords: Autophagy; Beclin-1; Caspase-3; LC3-II/I; rhACE2