title: Protective effect of microRNA-30b on hypoxia/reoxygenation-induced apoptosis in H9C2 cardiomyocytes.
abstract: We examined the protective role of microRNA-30b (miR-30b) in ischemia-reperfusion (I/R)-induced injury in rat H9C2 cardiomyocytes. H9C2 cells were subjected to hypoxia-reoxygenation (H/R) treatment to simulate ischemia-reperfusion (I/R) injury. H9C2 cells were divided into: vehicle control (VC) group; scrambled inhibitors (INC) group; scrambled mimics (MNC) group; H/R+VC group; H/R+INC group; H/R+mimics group. H/R induced apoptosis was detected by flow cytometry and the pathways involved in miR-30b-mediated protection were examined by analyzing the expression of miR-30b, Bcl-2, Bax, Caspase-3, KRAS, p-AKT and total AKT in H9C2 cells. Overexpression of miR-30b mimic (H/R+mimics group) significantly increased Bcl-2 and Bcl-2/Bax levels and decreased Bax and Caspase-3 levels, compared with the H/R+VC group (all P<0.05). Consistent with this, the apoptosis rate was significantly decreased in the H/R+mimics group (P<0.05) compared with the H/R+VC group. Western blot analysis revealed that overexpression of miR-30b mimic resulted in significantly increase in AKT activation and decreased KRAS, compared to the H/R+VC group (both P<0.05). In conclusion, the H/R induced apoptosis decreased miR-30b expression, but over-expression of miR-30b inhibited H/R induced apoptosis. The observed miR-30b-mediated protection against H/R induced apoptosis involved the upregulation of Ras-PI3K-Akt pathway.
authors.affiliation: Department of Emergency, The First Affiliated Hospital, Zhejiang University, Hangzhou 310003, China.
authors.forename: Tong
authors.initials: T
authors.lastname: Li
authors.affiliation: Department of Cardiology, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Ze-Lin
authors.initials: ZL
authors.lastname: Sun
authors.affiliation: Department of Cardiology, Xiangya Hospital, Central South University, Changsha 410008, China. Electronic address: xieqiying725@126.com.
authors.forename: Qi Ying
authors.initials: QY
authors.lastname: Xie
mesh_terms: D000818:Animals; D017209:Apoptosis; D015687:Cell Hypoxia; D002460:Cell Line; D005786:Gene Expression Regulation; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction
chemical_list: C556157:MIRN30 microRNA, rat; D035683:MicroRNAs; D051057:Proto-Oncogene Proteins c-akt
keywords: Cell apoptosis; H9C2 cardiomyocytes; Hypoxia–reoxygenation; Ischemia–reperfusion; MicroRNA-30b; Ras–PI3K–Akt