MiR-27b attenuates mitochondrial oxidative stress and inflammation in endothelial cells

Prattichizzo, Francesco; Martino, Elisa; Anastasio, Camilla; Mele, Luigi; Grotta, Rosalba La; Sardu, Celestino; Ceriello, Antonio; Marfella, Raffaele; Paolisso, Giuseppe; Luisa, Balestrieri Maria

doi:10.1016/j.redox.2023.102681

Cited by 19 publications

(30 citation statements)

References 53 publications

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“…Here, ATF4 was overexpressed in response to static vein culture, which may drive CSE overexpression in that condition. The micro RNA miR-27b, which is highly expressed in EC and involved in angiogenesis [ 45 , 46 ], has been described to down-regulate CTH expression [ 20 ]. However, this mechanism has been described only in EC and miR-27b is down-regulated in EC in pathological condition [ 20 ].…”

Section: Discussionmentioning

confidence: 99%

Cystathionine Gamma Lyase Is Regulated by Flow and Controls Smooth Muscle Migration in Human Saphenous Vein

Zhao,

Deslarzes-Dubuis,

Urfer

et al. 2023

Antioxidants

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The saphenous vein is the conduit of choice for bypass grafting. Unfortunately, the hemodynamic stress associated with the arterial environment of the bypass vein graft leads to the development of intimal hyperplasia (IH), an excessive cellular growth and collagen deposition that results in restenosis and secondary graft occlusion. Hydrogen sulfide (H2S) is a ubiquitous redox-modifying gasotransmitter that inhibits IH. H2S is produced via the reverse trans-sulfuration pathway by three enzymes: cystathionine γ-lyase (CSE), cystathionine β-synthase (CBS) and 3-mercaptopyruvate sulfurtransferase (3-MST). However, the expression and regulation of these enzymes in the human vasculature remains unclear. Here, we investigated the expression of CSE, CBS and 3-MST in segments of native human saphenous vein and large arteries. Furthermore, we evaluated the regulation of these enzymes in vein segments cultured under static, venous (7 mmHg pressure) or arterial (100 mmHg pressure) pressure. CSE was expressed in the media, neointima and intima of the vessels and was negatively regulated by arterial shear stress. Adenoviral-mediated CSE overexpression or RNA interference-mediated CSE knock-down revealed that CSE inhibited primary human VSMC migration but not proliferation. We propose that high shear stress in arteriovenous bypass grafts inhibits CSE expression in both the media and endothelium, which may contribute to increased VSMC migration in the context of IH.

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Section: Discussionmentioning

confidence: 99%

Cystathionine Gamma Lyase Is Regulated by Flow and Controls Smooth Muscle Migration in Human Saphenous Vein

Zhao,

Deslarzes-Dubuis,

Urfer

et al. 2023

Antioxidants

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“…Protein lysis, homogenate resolving by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), nitrocellulose membrane transfer, and chemiluminescent analysis were performed as previously described [ 42 , 43 ]. The following primary antibodies were used for western blotting detection: anti-sodium-glucose cotransporter-2 (SGLT2, 1:500, Abcam, Cambridge, UK), anti-sirtuin 3 (SIRT3, 1:500, Cell Signaling Technology, Danvers, MA, USA), anti-dipeptidyl peptidase-4 (DPP4, 1:500, Abcam, Cambridge, UK) anti-5' AMP-activated protein kinase alpha-1 (AMPK, 1:1000, Invitrogen, Waltham, Ma, USA), anti-phospho-AMPK (1:1000, Invitrogen, Waltham, Ma, USA), anti-C/EBP homologous protein (CHOP, 1:1000, Elabscience Biotechnology Inc., Houston, TX, USA), anti-protein kinase RNA-like ER kinase (PERK, 1:1000, Elabscience Biotechnology Inc., Houston, TX, USA), anti-activating transcription factor 6 (ATF6, 1:1000, Biorbyt, Cambridge, UK), anti-microtubule-associated protein 1 light chain 3B (LC3B, 1:1000, Abcam, Cambridge, UK), anti-actin (1:5000, Abcam, Cambridge, UK), and anti-α-tubulin (1:3000, Elabscience Biotechnology Inc., Houston, TX, USA).…”

Section: Methodsmentioning

confidence: 99%

SGLT2 inhibitor promotes mitochondrial dysfunction and ER-phagy in colorectal cancer cells

Anastasio,

Donisi,

Del Vecchio

et al. 2024

Cell Mol Biol Lett

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Background Sodium-glucose transporter 2 (SGLT2) inhibitors (iSGLT2) are approved medications for type 2 diabetes. Recent studies indicate that iSGLT2 inhibit the growth of some cancer cells. However, the mechanism(s) remains to be fully elucidated. Methods The SGLT2 levels were determined in normal colon CCD 841 CoN and, HCT 116, HT-29, SW480 and LoVo colorectal cancer (CRC) cell lines by quantitative real-time PCR and western blot. The effect of iSGLT2 canagliflozin on cell proliferation was examined using CCK-8, as its role on CRC cells metabolism and tumorigenesis has been evaluated by XF HS Seahorse Bioanalyzer and flow cytometric analyses. Transient gene silencing experiments and analysis of protein–protein interaction network were conducted to evaluate the SGLT2 molecular targets in CRC cells. Results Data showed that the treatment with iSGLT2 (50 µM) for 72 h induced cell cycle arrest (p < 0.001), impaired glucose and energetic metabolism (p < 0.001), promoted apoptotic cell death and ER stress flowing into autophagy (p < 0.001) in HCT 116 and HT-29 cells. These cellular events were accompanied by sirtuin 3 (SIRT3) upregulation (p < 0.01), as also supported by SIRT3 transient silencing experiments resulting in the attenuation of the effects of iSGLT2 on the cellular metabolic/energetic alterations and the induction of programmed cell death. The identification and validation of dipeptidyl peptidase 4 (DPP4) as potential common target of SGLT2 and SIRT3 were also assessed. Conclusions These results deepened knowledge on the iSGLT2 contribution in limiting CRC tumorigenesis unveiling the SGLT2/SIRT3 axis in the cytotoxic mechanisms. Graphical Abstract

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“…EC from human aorta (TeloHAEC, CRL-4052, ATCC, Manassas, VA, USA) and primary human umbilical vein (HUVEC, PCS-100-010, ATCC, Manassas, VA, USA) were grown as previously reported [ 50 ]. To mimic septic inflammatory condition, EC were treated up to 24 h with increasing concentrations (0.5–10 µg/mL) of lipopolysaccharides (LPS, L5543, Sigma-Aldrich, St. Louis, MO, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Cell viability and cytotoxicity assays were assessed as previously described [ 11 , 50 ], following the instructions of the manufacturer. The lactate dehydrogenase (LDH) release (%) in culture media was used to determine cytotoxicity.…”

Section: Methodsmentioning

confidence: 99%

MiR-15b-5p and PCSK9 inhibition reduces lipopolysaccharide-induced endothelial dysfunction by targeting SIRT4

et al. 2023

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Background Endothelial dysfunction and deregulated microRNAs (miRNAs) participate in the development of sepsis and are associated with septic organ failure and death. Here, we explored the role of miR-15b-5p on inflammatory pathways in lipopolysaccharide (LPS)-treated human endothelial cells, HUVEC and TeloHAEC. Methods The miR-15b-5p levels were evaluated in LPS-stimulated HUVEC and TeloHAEC cells by quantitative real-time PCR (qRT–PCR). Functional experiments using cell counting kit-8 (CCK-8), transfection with antagomir, and enzyme-linked immunosorbent assays (ELISA) were conducted, along with investigation of pyroptosis, apoptosis, autophagy, and mitochondrial reactive oxygen species (ROS) by cytofluorometric analysis and verified by fluorescence microscopy. Sirtuin 4 (SIRT4) levels were detected by ELISA and immunoblotting, while proprotein convertase subtilisin-kexin type 9 (PCSK9) expression was determined by flow cytometry (FACS) and immunofluorescence analyses. Dual-luciferase reporter evaluation was performed to confirm the miR-15b-5p–SIRT4 interaction. Results The results showed a correlation among miR-15b-5p, PCSK9, and SIRT4 levels in septic HUVEC and TeloHAEC. Inhibition of miR-15b-5p upregulated SIRT4 content, alleviated sepsis-related inflammatory pathways, attenuated mitochondrial stress, and prevented apoptosis, pyroptosis, and autophagic mechanisms. Finally, a PCSK9 inhibitor (i-PCSK9) was used to analyze the involvement of PCSK9 in septic endothelial injury. i-PCSK9 treatment increased SIRT4 protein levels, opposed the septic inflammatory cascade leading to pyroptosis and autophagy, and strengthened the protective role of miR-15b-5p inhibition. Increased luciferase signal validated the miR-15b-5p–SIRT4 binding. Conclusions Our in vitro findings suggested the miR-15b-5p–SIRT4 axis as a suitable target for LPS-induced inflammatory pathways occurring in sepsis, and provide additional knowledge on the beneficial effect of i-PCSK9 in preventing vascular damage by targeting SIRT4. Graphical Abstract

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MiR-27b attenuates mitochondrial oxidative stress and inflammation in endothelial cells

Cited by 19 publications

References 53 publications

Cystathionine Gamma Lyase Is Regulated by Flow and Controls Smooth Muscle Migration in Human Saphenous Vein

Cystathionine Gamma Lyase Is Regulated by Flow and Controls Smooth Muscle Migration in Human Saphenous Vein

SGLT2 inhibitor promotes mitochondrial dysfunction and ER-phagy in colorectal cancer cells

MiR-15b-5p and PCSK9 inhibition reduces lipopolysaccharide-induced endothelial dysfunction by targeting SIRT4

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