LncRNA FGD5-AS1 Targeting miR-16-5p/CREB1 Axis Alleviates Hypoxia/Reoxygenation Induced Apoptosis of Rat H9c2 Cardiomyocytes
Objective:To explore the effect of long non-coding RNA(LncRNA)FGD5 antisense RNA1(FGD5-AS1)on the apoptosis of rat H9c2 cardiomyocytes induced by hypoxia/reoxygenation(H/R)and the regulatory mechanism of microRNA-16-5p/cAMP response element-binding protein 1(miR-16-5p/CREB1)axis.Methods:H9c2 cardiomyocytes were divided into control group and H/R group(hypoxia for 6 h,reoxygenation for 6 h).H/R group cells were transfected separately,then divided into oe-NC group,oe-FGD5-AS1 group,oe-FGD5-AS1+miR-16-5p mimic-NC group,and oe-FGD5-AS1+miR-16-5p mimic group.Real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used to detect the mRNA levels of FGD5-AS1,miR-16-5p and CREB1 in cells.The expression of cleaved Caspase-3,B-cell lymphoma/leukemia-2(Bcl-2)and Bcl-2 associated X protein(Bax)were measured by Western Blot.Cell survival rate was measured by CCK-8 method.The apoptosis rate was detected by flow cytometry.The targeting relationship between miR-16-5p mRNA,FGD5-AS1 and CREB1 was verified by double luciferase activity assay.Results:Compared with the control group,mRNA levels of FGD5-AS1 and CREB1,cell survival rate,and Bcl-2 protein expression in H/R group decreased(P<0.05),miR-16-5p mRNA levels,apoptosis rate,and cleaved Caspase-3 and Bax protein expressions increased(P<0.05).Compared with H/R group and oe-NC group,mRNA levels of FGD5-AS1 and CREB1,cell survival rate,Bcl-2 protein expression increased in H9c2 cells transfected with FGD5-AS1 gene overexpression,the apoptosis rate,levels of miR-16-5p,cleaved Caspase-3,and Bax decreased(P<0.05).Dual luciferase activity test verified that FGD5-AS1 and CREB1 had binding sites with miR-16-5p.Conclusion:Overexpression of FGD5-AS1 may inhibit H/R-induced apoptosis of H9c2 cardiomyocytes through targeted down-regulation of miR-16-5p expression and up-regulation of CREB1 expression.
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