Folic Acid Alleviates High Glucose and Fat-Induced Pyroptosis via Inhibition of the Hippo Signal Pathway on H9C2 Cells

Abstract

Diabetic cardiomyopathy (DCM) is the leading cause of death in diabetic patients. Folic acid has a protective effect on diabetes-induced cardiomyocyte damage. The aim of this study was to explore the effects of folic acid on cardiomyocytes cultured under high glucose and fat (HGF) conditions and type 2 diabetes mellitus (T2DM) mice, and elucidate the underlying mechanisms. Bioinformatics analysis was used to identify the potential drugs through the Drug-Gene Interaction database. H9C2 cardiomyocytes were cultured with 30 mM glucose and 500 nM palmitic acid in the presence or absence of folic acid or YAP1 inhibitor (verteporfin) or YAP1 siRNA. The cell viability and lactate dehydrogenase (LDH) release were measured using specific assay kits. Pyroptosis was detected by flow cytometry. The concentrations of IL-1β and IL-18 in the supernatants were measured by ELISA. The NLRP3, ASC and caspase-1 mRNA levels were detected by qRT-PCR and that the proteins expression of NLRP3, ASC, cleaved caspase-1 (p10), caspase-1, YAP1, p-YAP1, LATS1 and P-LATS1 were detected by Western blotting. C57BL/6 mice were fed with high fat diet (HFD) combined with streptozotocin (STZ) intraperitoneally to establish a T2DM model, folic acid or PBS treatment for 8 weeks by oral gavage, blood glucose and body weight were measured every 4 weeks, mouse heart tissue was used to detect pyroptosis and hippo signaling pathway related protein expression. We identified 427 differentially expressed genes in the cardiac tissues of high fat diet + streptozotocin mice, among the 30 most significantly DEGs, folic acid was predicted to be the most likely therapeutic drug. Folic acid alleviated HGF-induced cell damage in vitro and in vivo by decreasing activation of the Hippo pathway, as indicated by lower LDH release and increased cell viability, and decreased expression of NLRP3, ASC, cleaved caspase-1, IL-1β, IL-18, p-YAP and p-LATS. Verteporfin or YAP1 siRNA neutralized the protective effect of folic acid by reversing YAP1-induced pyroptosis. Folic acid reduced NLRP3 inflammasome-mediated pyroptosis by down-regulating the Hippo signaling pathway, thereby effectively reducing T2DM-induced damage in H9C2 cells and animals.

Oncogenic role of a long-term high-fat diet-induced DNM1 in hepatocellular carcinoma

“…Down-regulating the hippo signaling pathway effectively reduced high fat diet and streptozotocin induced damage in H9C2 cardiomyocytes and mice. Folic acid was found to alleviate high glucose and high fatinduced cell death triggered by proin ammatory signals via the inhibition of the hippo signal pathway [27] . Data from our current study supported the postulation that DNM1 and /or hippo signaling pathways could serve as therapeutic targets for high fat associated in ammation that potentiates HCC risk.…”

Different exercise training intensities prevent type 2 diabetes mellitus-induced myocardial injury in male mice

“…To establish the T2DM model, 50 mg/kg streptozotocin (sigma, Germany) was injected into the enterocoelia of mice in the experimental groups, and mice were fed a HFD (fat 60%, protein 20%, carbohydrate 20%, Greisway Biotechnology Co., Ltd. Suzhou, China). 53 Experimental group mice were fed high-fat foods for six weeks. Subsequently, mice were divided into the two exercise training groups and subjected to exercise five days per week for 24 consecutive weeks.…”

Potential clinical biomarkers and perspectives in diabetic cardiomyopathy

“…Tilianin and syringin were used together by Yao et al to improve cardiac function in diabetic rats by lowering levels of diabetes/hyperglycemia-induced oxidative stress and regulating the TLR4/NF-κB/NLRP3 and PGC1/SIRT3/mitochondrial pathways [ 49 ]. Folic acid slows the development of DCM by blocking NLRP3-mediated inflammatory vesicle-mediated cellular scorching through downregulation of the Hippo signaling pathway [ 50 ]. Inhibiting TGF-β1/Smad signaling and lowering NLRP3 inflammasome activation, the extract of Coriolus Versicolor protects against DCM in rats by decreasing myocardial fibrosis and inflammation [ 51 ].…”