Knockdown of miR-372-3p Inhibits the Development of Diabetic Cardiomyopathy by Accelerating Angiogenesis via Activating the PI3K/AKT/mTOR/HIF-1α Signaling Pathway and Suppressing Oxidative Stress

Abstract

Background. DCM is the most common and malignant complication of diabetes. It is characterized by myocardial dilatation, hypertrophy, fibrosis, ventricular remodeling, and contractile dysfunction. Although many studies have demonstrated the function of miRNAs in the progression of DCM, but the specific role of miR-372-3p in DCM remains unknown. Methods. C57/BL6J mice were used to construct mouse models of DCM by intraperitoneal injection of STZ (50 mg/kg/d) for 5 consecutive days. Then the mice were randomly divided into model group (intramyocardial injection of empty lentivirus) and miR-372-3p KD group (intramyocardial injection of miR-372-3p KD lentivirus at 10 9 /mouse). Besides, the control group (injection of 0.9% normal saline) was also set up. LY294002, a PI3K inhibitor, was employed in the current study. Western blotting, immunofluorescence staining, quantitative ultrasound method, Masson's trichrome staining, and bioinformatics analysis were performed. Results. It was found that miR-372-3p KD significantly improved left ventricular dysfunction and cardiac hypertrophy in DCM mice. Furthermore, it also improved myocardial interstitial fibrosis and remodeling in DCM mice. Immunofluorescence staining and RT-qPCR revealed that miR-372-3p KD might accelerate cardiac remodeling by increasing angiogenesis in DCM mice. Western blotting results revealed that miR-372-3p was an upstream target of the PI3K/ AKT-mTOR and HIF-1α signals, as well as NOX2, NOX4, which were responsible for angiogenesis in DCM mice. Besides, the in vitro experiment showed that LY294002 markedly diminished the increased expression levels of p-PI3K, AKT, p-mTOR, p-P70S6K, HIF-1α, NOX2, and NOX4 in the model group and the miR-372-3p KD group, suggesting that PI3K signaling pathway and oxidative stress are involved in miR-372-3p KD-induced angiogenesis in HG-stimulated C166 cells. Conclusions. MiR-372-3p KD inhibits the development of DCM via activating the PI3K/AKT/mTOR/HIF-1α signaling pathway or suppressing oxidative stress. This offers an applicable biomarker for DCM treatment.

Enrichment of miR-17-5p enhances the protective effects of EPC-EXs on vascular and skeletal muscle injury in a diabetic hind limb ischemia model

“…Our findings demonstrate for the first time that EPC-EXs miR-17-5p protect ECs and C2C12 cells from Hypoxia plus HG injury by targeting SPRED1 and activating the PI3K/Akt pathway, which is consistent with a previous study showing that the SPRED1-PI3K/Akt pathway is involved in regulating ECs and muscle cell functions [ 11 , 24 ]. A PI3K inhibitor (LY294002) has been used to address the participation of the PI3K/Akt pathway in regulating multiple cell functions [ 50 , 51 ]. Using a PI3K inhibitor, we further demonstrated that EPC-EXs enriched with miR-17-5p ameliorated HG plus hypoxia-induced EC (increasing viability, migration, and tube formation) and C2C12 cell (increasing viability and myotube size) dysfunction and apoptosis by activating the PI3K/Akt pathway.…”

Single-cell RNA sequencing reveals aortic cellular heterogeneity in MSC_ITGB3-treated ApoE-/- mice on a high-fat diet

“…The PI3K-Akt signaling pathway and ECM-receptor interaction were significantly enriched in EC_1. PI3K-PKB/Akt is a highly conserved signaling pathway, and its activation mediates many cellular functions, including angiogenesis, survival, growth, transcription, proliferation, and apoptosis [31, 32]. ECMs interact with cell surface receptors to regulate cell behavior, such as cell communication, proliferation, migration, and adhesion [33, 34].…”

“…The PI3K-Akt signaling pathway and ECM-receptor interaction were significantly enriched in EC_1. PI3K-PKB/Akt is a highly conserved signaling pathway, and its activation mediates many cellular functions, including angiogenesis, survival, growth, transcription, proliferation, and apoptosis [31,32].…”

The Role of Epicardial Adipose Tissue-Derived MicroRNAs in the Regulation of Cardiovascular Disease: A Narrative Review

“…MicroRNA-327-3p was another EAT-derived miRNA increased in CAD patients [ 85 ], and it is involved in the development of diabetic cardiomyopathy (DCM). Recently, it was reported that the downregulation of miR-372-3p promotes angiogenesis while suppressing oxidative stress, presumably by targeting phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) in DCM mice [ 99 ]. That is to say that miR-372-3p can negatively regulate angiogenesis and increase oxidative stress.…”

“… MicroRNA Underlying CVD [Ref.] Expression Change Reported Target, Effects Relevant to Cardiovascular System Reference miR-135b 1 CAD [ 85 ] Increased 2 HIP1, inhibited 3 EC proliferation, migration, and tube formation [ 92 ] 4 GPX4, promoted ferroptosis of 5 CM [ 95 ] miR-1231 CAD [ 85 ] Increased 6 CACNA2D2, aggravated arrhythmia [ 97 ] miR-372-3p CAD [ 85 ] Increased 7 PIK3CA, downregulation in 8 DCM mice promoted angiogenesis while suppressing oxidative stress [ 99 ] miR-6870-3p CAD [ 100 ] Increased 9 TOLLIP, increased the expression of inflammatory genes ( 10 TLR4, 11 IL-6, 12 JNK, 13 NF-κB, and 14 TNF-α) [ 100 ] miR-34a 15 SCD [ 91 ] …”