The effect of long non-coding RNA maternal expression 3 on high glucose-induced islet β cell injury through TGF-β-Smad2/3 signaling pathway
Objective To investigate the effect of long-chain non-coding ribonucleic acid maternal expression 3(MEG3)on high glucose-induced pancreatic islet β-cell injury through the transforming growth factor-β-Smad 2/3 signaling pathway.Methods INS-1 rat insulinoma cells were divided into normal control(Con)group,HG group,negative control(si-NC)group,HG+si-NC group,siRNA targeting MEG3(si-MEG3)group,HG+si-MEG3 group,HG+si-MEG3+TGF-β-Smad2/3 signaling pathway inhibitor(HG+si-MEG3+LY2109761)group.Cell viability was detected by cell counting kit-8(CCK-8).The mRNA expression of MEG3 and pyroptosis-associated markers was detected by qRT-PCR.Autophagic vesicles were tested by transmission electron microscopy,and expression of autophagy-associated proteins and pyroptosis-associated proteins was detected by Western blot and immunofluorescence.TGF-β-Smad2/3 protein expression was detected by Western blot.Results Glucose downregulated MEG3 mRNA expression in a dose-dependent manner.Compared with si-NC group,p-Smad2,TGF-β1,Beclin1,LC3-II/LC3-I,NLRP3,GSDMD-N,IL-1β,ASC,and c-Caspase1 protein expression was elevated(P<0.05),and cell viability was decreased in HG+si-NC group(P<0.05).Compared with the HG+si-NC group,TGF-β1,NLRP3,GSDMD-N,IL-1β,ASC,c-Caspase1 protein expression was elevated(P<0.05),and p-Smad2,cell viability,Beclin1,and LC3-II/LC3-I were decreased in the HG+si-MEG3 group(P<0.05).Compared with HG+si-MEG3 group,NLRP3,GSDMD-N,IL-1β,ASC,and c-Caspase1 protein expression was elevated(P<0.05),and cell viability,p-Smad2,TGF-β1,Beclin1,and LC3-II/LC3-I protein expression was decreased in HG+si-MEG3+LY2109761 group(P<0.05).Conclusions MEG3 regulates autophagy and pyroptosis in pancreatic β-cells through the TGF-β-Smad2/3 signaling pathway.
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