LncRNA H19吸附miR-590-3p调控GDF-15对心肌细胞缺氧/复氧损伤减轻的作用研究OA

Objective:To analyze the effects of LncRNA H19 on GDF-15 regulation of hypoxia/reoxy-genation injury in cardiomyocytes by miR-590-3p.Methods:The H9c2 rat cardiomyocyte cell line was se-lected for cell culture and transfection.Cells were divided into the following groups:blank group subjected to routine culture without any treatment;hypoxia group subjected to hypoxia treatment only,cultured in 94%N2,5%CO2,1%O2 environment for 4h in DMEM medium without glucose or serum;model group subjected to hypoxia/reoxygenation treatment only;lncRNA H19 overexpression group subjected to hypoxia/reoxygenation treatment after pcDNA3.1-H19 transfection;miR-590-3p mimic group undergoing hypoxia/reoxygenation treatment after miR-590-3p mimic transfection;lncRNA H19 overexpression+miR-590-3p inhibitor group subjected to pcDNA3.1-H19 transfection first,followed by miR-590-3p inhibitor transfection after 24h,then hypoxia/reoxygenation treatment;negative control group(NC)subjected to hypoxia/reoxygenation treatment after pcDNA3.1-NC+miR-NC transfection.H19 overexpression+miR-590-3p mimic+GDF-15 overex-pression group subjected to pcDNA3.1-H19 transfection first,followed by co-transfection of miR-590-3p mimic and pcDNA3.1-GDF-15 after 24 h,then hypoxia/reoxygenation treatment.Meanwhile,animal experi-ments were also conducted in parallel.The viability of cardiomyocytes was calculated in each group;the per-centage of apoptotic cells was recordd;the expression levels of LncRNA H19,miR-590-3p,and GDF-15 mRNA were detected;the protein expression of GDF-15 was measured;and the levels of TNF-α,IL-6,LDH,and CK-MB were determined.The dual-luciferase reporter gene assay was used to validate the regula-tory relationship among LncRNA H19,miR-590-3p,and GDF-15.and myocardial tissue apoptosis,as well as to detect the phosphorylation levels of Smad2,Akt,and mTOR pathway proteins downstream of GDF-15.Results:Compared with the model group,the H19 overexpression group exhibited increased cell viability and reduced apoptosis rate,upregulated expression of LncRNA H19 and GDF-15,downregulated expression of miR-590-3p,and decreased levels of inflammatory factors and myocardial enzymes(P<0.05).The miR-590-3p mimic group showed the opposite trend(P<0.05).Compared with the H19-WT+miR-NC group,the H19-WT+miR-590-3p group demonstrated a reduced luciferase activity ratio(P<0.05).Similarly,the GDF-15-WT+miR-590-3p group showed a reduced luciferase activity ratio compared to the GDF-15-WT+miR-NC group(P<0.05).Functional recovery experiments demonstrated that the combination of H19 over-expression,miR-590-3p mimic,and GDF-15 overexpression significantly reversed the myocardial cell injury induced by miR-590-3p mimic,restored cellular viability,and inhibited apoptosis(P<0.05).Conclu-sions:By adsorbing miR-590-3p and relieving its inhibition on GDF-15,lncRNA H19 can alleviate myocar-dial cell hypoxia/reoxygenation injury,providing a new target for AMI treatment.