Epigenetic repression of THBD transcription by BRG1 contributes to deep vein thrombosis.

BACKGROUND: Deep vein thrombosis (DVT) with its major complication, pulmonary embolism, is a global health problem. Endothelial dysfunction is involved in the pathogenesis of DVT. We have previously demonstrated that endothelial specific deletion of Brahma-related gene 1 (BRG1) ameliorates atherosclerosis and aneurysm in animal models. Whether endothelial BRG1 contributes to DVT development remains undetermined. METHODS: DVT was induced in mice by ligation of inferior vena cava. Deletion of BRG1 in endothelial cells was achieved by crossing the Cdh5-ERT-Cre mice with the Brg1loxp/loxp mice. RESULTS: Here we report that compared to the wild type mice, BRG1 conditional knockout (CKO) mice displayed substantially decreased DVT susceptibility characterized by decreased weight and size of thrombus and reduced immune infiltration. In endothelial cells, thrombomodulin (THBD) expression was significantly decreased by TNF-α stimulation, while BRG1 knockdown or inhibition recovered THBD expression. Further analysis revealed that BRG1 deficiency decreased the CpG methylation levels of the THBD promoter induced by TNF-α. Mechanistically, BRG1 directly upregulated DNMT1 expression after TNF-α treatment in endothelial cells. More importantly, administration of a small-molecule BRG1 inhibitor PFI-3 displayed potent preventive and therapeutic potentials in the DVT model. CONCLUSIONS: Our findings implicate BRG1 as an important regulator of DVT pathogenesis likely through epigenetic regulation of THBD expression in endothelial cells and provide translational proof-of-concept for targeting BRG1 in DVT intervention.

Oenothein B boosts antioxidant capacity and supports metabolic pathways that regulate antioxidant defense in Caenorhabditis elegans.

Oenothein B (OEB) has various biological functions, although few studies have focused on its effect on in vivo metabolic phenotypes. In the present study, the systematic antioxidant activity of OEB was evaluated both in vitro and in vivo, and the effect of OEB on metabolic pathways related to antioxidant capacity of Caenorhabditis elegans (C. elegans) was explored. Our findings indicate that OEB exhibits great antioxidant capacity and ability to scavenge free radicals and that OEB treatment can protect RAW 264.7 macrophages from oxidative damage by increasing superoxide dismutase (SOD) activity, catalase (CAT) activity and glutathione (GSH) content and the corresponding gene expression (sod2, cat, gpx1), while decreasing malonic dialdehyde (MDA) content. Moreover, OEB treatment significantly reduced ROS accumulation under oxidative stress conditions and increased glutathione peroxidase (GPx) activity and decreased MDA content in C. elegans. Metabolomics analysis revealed that sixteen out of forty-two significantly altered metabolites were selected as potential biomarkers related to alterations in the antioxidant status of worms, including metabolic pathways involved in amino acid metabolism, taurine and hypotaurine metabolism, lipid metabolism, and purine metabolism. Overall, our results provide new insights into the effects of OEB treatment on antioxidant capacity and metabolism that suggest that OEB could be a potentially good source of natural antioxidants.

Ginkgolide K attenuates neuronal injury after ischemic stroke by inhibiting mitochondrial fission and GSK-3ß-dependent increases in mitochondrial membrane permeability.

Ginkgolide K (GK) belongs to the ginkgolide family of natural compounds found in Ginkgo biloba leaves, which have been used for centuries to treat cerebrovascular and cardiovascular diseases. We evaluated the protective effects of GK against neuronal apoptosis by assessing its ability to sustain mitochondrial integrity and function. Co-immunoprecipitation showed that Drp1 binding to GSK-3ß was increased after an oxygen-glucose deprivation/reperfusion (OGD/R) insult in cultured neuroblastoma cells. This induced Drp1 and GSK-3ß translocation to mitochondria and mitochondrial dysfunction, which was attenuated by GK. GK also reduced mitochondrial fission by increasing Drp1 phosphorylation at Ser637 and inhibiting mitochondrial Drp1 recruitment. In addition, GK exposure induced GSK-3ß phosphorylation at Ser9 and enhanced the interaction between adenine nucleotide translocator (ANT) and p-GSK-3ß. This interaction suppressed the interaction between ANT and cyclophilin D (CypD), which inhibited mitochondrial permeability transition pore (mPTP) opening. Similarly, suppression of mitochondrial fission by Mdivi-1 also inhibited GSK-3ß-induced mPTP opening. Treating mice with GK prevented GSK-3ß and Drp1 translocation to mitochondria and attenuated mitochondrial dysfunction after middle cerebral artery occlusion. We therefore propose that by inhibiting mitochondrial fission and attenuating mPTP opening, GK exerts neuroprotective effects that mitigate or prevent neuronal damage secondary to ischemic stroke.