title: miR-30b inhibits autophagy to alleviate hepatic ischemia-reperfusion injury via decreasing the Atg12-Atg5 conjugate.
abstract: OBJECTIVE
  To explore the role and potential mechanism of miR-30b regulation of autophagy in hepatic ischemia-reperfusion injury (IRI).
  
  
  METHODS
  An animal model of hepatic IRI was generated in C57BL/6 mice. For in vitro studies, AML12 cells were immersed in mineral oil for 1 h and then cultured in complete Dulbecco's Modified Eagle's Medium (DMEM)/F12 to simulate IRI. Mice and cells were transfected with miR-30b agomir/mimics or antagomir/inhibitor to examine the effect of miR-30b on autophagy to promote hepatic IRI. The expression of miR-30b was measured by real-time polymerase chain reaction. Apoptotic cells were detected by terminal uridine nick-end labeling (TUNEL) staining, and cell viability was detected by methylthiazole tetrazolium assay. The expression of light chain 3, autophagy-related gene (Atg)12, Atg5, P62, and caspase-3 were detected by western blotting analysis.
  
  
  RESULTS
  miR-30b levels were significantly downregulated after hepatic IRI, and the numbers of autophagosomes were increased in response to IRI both in vivo and in vitro. These findings demonstrate that low levels of miR-30b could promote hepatic IRI. Furthermore, we found that miR-30b interacted with Atg12-Atg5 conjugate by binding to Atg12. Overexpression of miR-30b diminished Atg12 and Atg12-Atg5 conjugate levels, which promoted autophagy in response to IR. In contrast, downregulation of miR-30b was associated with increased Atg12-Atg5 conjugate levels and increased autophagy.
  
  
  CONCLUSIONS
  miR-30b inhibited autophagy to alleviate hepatic ischemia-reperfusion injury via decreasing the Atg12-Atg5 conjugate.
authors.affiliation: Shi-Peng Li, Jin-Dan He, Zhen Wang, Yao Yu, Hai-Ming Zhang, Jian-Jun Zhang, Zhong-Yang Shen, First Central Clinical College of Tianjin Medical University, Tianjin 300192, China.
authors.forename: Shi-Peng
authors.initials: SP
authors.lastname: Li
authors.affiliation: Shi-Peng Li, Jin-Dan He, Zhen Wang, Yao Yu, Hai-Ming Zhang, Jian-Jun Zhang, Zhong-Yang Shen, First Central Clinical College of Tianjin Medical University, Tianjin 300192, China.
authors.forename: Jin-Dan
authors.initials: JD
authors.lastname: He
authors.affiliation: Shi-Peng Li, Jin-Dan He, Zhen Wang, Yao Yu, Hai-Ming Zhang, Jian-Jun Zhang, Zhong-Yang Shen, First Central Clinical College of Tianjin Medical University, Tianjin 300192, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Shi-Peng Li, Jin-Dan He, Zhen Wang, Yao Yu, Hai-Ming Zhang, Jian-Jun Zhang, Zhong-Yang Shen, First Central Clinical College of Tianjin Medical University, Tianjin 300192, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Shi-Peng Li, Jin-Dan He, Zhen Wang, Yao Yu, Hai-Ming Zhang, Jian-Jun Zhang, Zhong-Yang Shen, First Central Clinical College of Tianjin Medical University, Tianjin 300192, China.
authors.forename: Shu-Yu
authors.initials: SY
authors.lastname: Fu
authors.affiliation: Shi-Peng Li, Jin-Dan He, Zhen Wang, Yao Yu, Hai-Ming Zhang, Jian-Jun Zhang, Zhong-Yang Shen, First Central Clinical College of Tianjin Medical University, Tianjin 300192, China.
authors.forename: Hai-Ming
authors.initials: HM
authors.lastname: Zhang
authors.affiliation: Shi-Peng Li, Jin-Dan He, Zhen Wang, Yao Yu, Hai-Ming Zhang, Jian-Jun Zhang, Zhong-Yang Shen, First Central Clinical College of Tianjin Medical University, Tianjin 300192, China.
authors.forename: Jian-Jun
authors.initials: JJ
authors.lastname: Zhang
authors.affiliation: Shi-Peng Li, Jin-Dan He, Zhen Wang, Yao Yu, Hai-Ming Zhang, Jian-Jun Zhang, Zhong-Yang Shen, First Central Clinical College of Tianjin Medical University, Tianjin 300192, China.
authors.forename: Zhong-Yang
authors.initials: ZY
authors.lastname: Shen
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D017209:Apoptosis; D001343:Autophagy; D000071192:Autophagy-Related Protein 12 / Q000235:genetics / Q000378:metabolism*; D000071187:Autophagy-Related Protein 5 / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002460:Cell Line; D004195:Disease Models, Animal; D015536:Down-Regulation; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008107:Liver Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D008297:Male; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015427:Reperfusion Injury / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D015398:Signal Transduction; D014162:Transfection
chemical_list: D020413:3' Untranslated Regions; C466624:ATG12 protein, human; C495773:Atg5 protein, mouse; D000071192:Autophagy-Related Protein 12; D000071187:Autophagy-Related Protein 5; D035683:MicroRNAs; C534715:Mirn30d microRNA, mouse; D012333:RNA, Messenger
keywords: Atg12-Atg5 conjugate; Autophagy; Hepatic ischemia-reperfusion injury; miR-30b