MiR-6321 regulates N6-methyladenosine methylation modification to alleviate myocardial ischemia-reperfusion injury in rats
LING Yuya
XU Tongtong
LYU Xiangwei
ZHAO Weikun
QIN Qiuyu
LI Yuechang
LU Yufen
Abstract：Objective To explore the effect of microRNA-6321(miR-6321)on myocardial ischemia-reperfusion injury(MIRI)in rats and further analyze the relationship between its mechanism of action and the methylation of N6-methyladenosine.Methods Twelve male Sprague-Dawley(SD)rats were randomly selected from 36 rats and divided into a pre-experimental sham operation group and a pre-experimental model group,with 6 rats in each group,to verify the MIRI model.In the pre-experimental sham operation group,the thread was inserted below the left anterior descending branch without ligation.In the pre-experimental model group,the ligation thread was removed in 30 min after ligation followed by reperfusion for 2 h.Subsequently,the remaining 24 rats were randomly divided into Sham operation group,model group(MIRI group),adeno-associated virus negative control group(AAV-NC group)and miR-6321 overexpression intervention group(AAV-miR-6321 group),with 6 rats in each group.The AAV-NC group and the AAV-miR-6321 group received a single injection of AAV-NC and AAV-miR-6321,respectively via the tail vein 2 weeks before modeling.The MIRI model in rats was established by ligation of the left anterior descending coronary artery.HE staining was used to observe the severity of myocardial tissue injury.Western blotting was employed to detect the expression of methyltransferase-like protein 3(METTL3),alkylation repair homolog 5(ALKBH5)and YTH domain family protein 1(YTHDF1)in myocardial tissue.Results Compared with the pre-experimental sham operation group,the expression level of ALKBH5 was significantly decreased,while those of METTL3 and YTHDF1 were obviously up-regulated in the pre-experimental model group(P<0.05).When compared with the sham operation group,the MIRI group exhibited more severe myocardial injury,decreased expression of miR-6321,elevated expression of METTL3 and YTHDF1(0.96±0.06 vs 0.52±0.08,1.04±0.04 vs 0.47±0.04,P<0.05),and reduced expression of ALKBH5(0.36±0.14 vs 0.89±0.09,P<0.05).AAV-miR-6321 injection resulted in significantly alleviated myocardial injury,enhanced expression of miR-6321,declined expression of METTL3 and YTHDF1(0.58±0.05 vs 0.96±0.06,0.55±0.04 vs 1.04±0.04,P<0.05),and increased expression of ALKBH5(0.76±0.14 vs 0.36±0.14,P<0.05)when compared with the MIRI group.Conclusion MiR-6321 can regulate N6-methyladenosine methylation modification to alleviate MIRI in rats.
Keywords：myocardial reperfusion injurymethylationmicroRNA-6321
Publication Date：2026-01-15
Online Publishing Date：2026-01-16(First online date of this platform, not the publication date of the document)
Pages：5( 102-106 )