Metformin activates autophagy and inhibits apoptosis and fetal heart abnormalities via the Dnmt3l/Usp18/STAT1 pathway.

Abstract

Gestational diabetes mellitus (GDM) is an important risk factor for developing congenital heart disease (CHD). It is urgent to find therapeutic drugs that reduce blood glucose and protect patients from CHD. In this study, rat models of type 1 (intraperitoneal injection with 50 mg/kg streptozocin) and type 2 (high fat/high sugar diet for 5 weeks plus 28 mg/kg streptozocin) gestational diabetes were established. Metformin (MET) had more beneficial effects on reducing maternal blood glucose levels and increasing the number of fetuses in the type 2 model than the type 1 model. The type 2 model was chosen for further study. Histological, apoptosis and western blotting analyses showed that MET activated autophagy and reduced apoptosis of cardiomyocytes in diabetic rats. After RNA-sequencing, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses, Dnmt3l was selected as a potential effector of MET. Western blotting, CCK8 and Hoechst 33342 staining analyses showed that Dnmt3l played an important role in the effects of MET on activating autophagy and inhibiting the excessive apoptosis induced by high glucose. 2-NP (STAT1 activator) counteracted the effects of MET. Further studies showed that Dnmt3l mediated the actions of MET by regulating STAT1(Y701) phosphorylation, and this process also required Usp18, which had decreased expression in fetal heart tissues exposed to hyperglycemia. These results demonstrate the effect of MET on reducing maternal blood glucose, reveal its new role in protecting fetal heart abnormalities via the Dnmt3l/Usp18/STAT1 pathway, and provide a theoretical basis for the treatment of MET on GDM and CHD.