Retinal thickness changes in different subfields reflect the volume change of cerebral white matter hyperintensity.

Purpose: To investigate the relationship between the retinal thickness in different subfields and the volume of white matter hyperintensity (WMH), with the hope to provide new evidence for the potential association between the retina and the brain. Methods: A total of 185 participants aged over 40 years were included in our study. Magnetic resonance imaging (MRI) was used to image the WMH, and WMH volume was quantitatively measured by a specific toolbox. The thickness of the total retina, the retinal nerve fiber layer (RNFL), and the ganglion cell and inner plexiform layer (GCIP) was measured by optical coherence tomography (OCT) in nine subfields. The association between retinal thickness and WMH volume was demonstrated using binary logistic regression and Pearson correlation analysis. Results: Participants were divided into two groups by the WMH volume (‰, standardized WMH volume) median. In the quartile-stratified binary logistic regression analysis, we found that the risk of higher WMH volume showed a positive linear trend correlation with the thickness of total retina (95% CI: 0.848 to 7.034; P for trend = 0.044)/ GCIP (95% CI: 1.263 to 10.549; P for trend = 0.038) at the central fovea, and a negative linear trend correlation with the thickness of nasal inner RNFL (95% CI: 0.086 to 0.787; P for trend = 0.012), nasal outer RNFL (95% CI: 0.058 to 0.561; P for trend = 0.004), and inferior outer RNFL (95% CI: 0.081 to 0.667; P for trend = 0.004), after adjusting for possible confounders. Correlation analysis results showed that WMH volume had a significant negative correlation with superior outer RNFL thickness (r = -0.171, P = 0.02) and nasal outer RNFL thickness (r = -0.208, P = 0.004). Conclusion: It is suggested that central fovea and outer retina thickness are respectively associated with WMH volume. OCT may be a biological marker for early detection and longitudinal monitoring of WMH.

Sirt6 inhibits vascular endothelial cell pyroptosis by regulation of the Lin28b/let-7 pathway in atherosclerosis.

Our previous study showed that Sirtuin 6 (Sirt6) plays an important role in the regulation of vascular endothelial cell inflammation. Recently, studies have reported that the RNA binding protein Lin28b directly regulates the let-7 microRNA (miRNA), which participates in the process of atherosclerosis (AS) by regulating inflammation. Pyroptosis is a form of programmed cell death that is accompanied by inflammation and is critical for AS. Thus, this study aimed to investigate the role of Sirt6 and Lin28b in vascular endothelial cell pyroptosis and the related mechanism. The present study showed that Lin28b expression was upregulated in the aortic intima and aorta of apolipoprotein E knockout (ApoE-/-) mice fed with a high-fat diet (HFD) for 8 or 12 weeks. Then, in vitro study found Lin28b was involved in tumor necrosis factor-α (TNF-α)-induced vascular endothelial cell pyroptosis, as indicated by the increased number of PI-positive cells and gasdermin D (GSDMD) cleavage, as well as the increased release of lactate dehydrogenase (LDH) and interleukin (IL)-1ß. Further studies demonstrated that TNF-α significantly decreased the expression of let-7, while Lin28b knockdown significantly increased the expression of let-7a, let-7d and let-7g. In addition, Sirt6 overexpression decreased Lin28b expression. Moreover, Sirt6 overexpression suppressed pyroptosis by decreasing the number of PI-positive cells and GSDMD cleavage, as well as by decreasing the release of LDH and IL-1ß in TNF-α-induced vascular endothelial cells. Further mechanistic studies revealed that Sirt6 directly interacted with and deacetylated Lin28b. Taken together, these findings indicate that Sirt6 inhibits vascular endothelial cell pyroptosis by negatively regulating the Lin28b/let-7 pathway in AS.

HDAC11 promotes both NLRP3/caspase-1/GSDMD and caspase-3/GSDME pathways causing pyroptosis via ERG in vascular endothelial cells.

Histone deacetylase 11 (HDAC11), a sole member of the class IV HDAC subfamily, participates in various cardiovascular diseases. Recent evidence showed that pyroptosis was a form of inflammatory programmed cell death and is critical for atherosclerosis (AS). However, little is known about the effect of HDAC11 on endothelial cell pyroptosis in AS. Thus, this study aims to investigate the role of HDAC11 in vascular endothelial cell pyroptosis and its molecular mechanism. Firstly, we found that HDAC11 expression was up-regulated and pyroptosis occurred in the aorta of ApoE-/- mice fed with a high-fat diet (HFD) for 8 or 12 weeks. Then, in vitro study found the treatment of human umbilical vein endothelial cells (HUVECs) with tumor necrosis factor-α (TNF-α) resulted in pyroptosis, as evidenced by activation of caspase-1 and caspase-3 activation, cleavage of downstream gasdermin D (GSDMD) and gasdermin E (GSDME/DFNA5), the release of pro-inflammatory cytokines interleukin (IL)-1ß, IL-6 and IL-18, as well as elevation of LDH activity and increase of propidium iodide (PI)-positive cells. Besides, TNF-α increased HDAC11 expression and induced pyroptosis via TNFR1 in HUVECs. HDAC11 knockdown mitigated pyroptosis by suppressing both NLRP3/caspase-1/GSDMD and caspase-3/GSDME pathways in TNF-α-induced HUVECs. Moreover, GSDME knockdown by siRNA significantly decreased pyroptosis and inflammatory response, while treatment with disulfiram or necrosulfonamide (NSA) further augmented the inhibitory effects of GSDME siRNA on pyroptosis and inflammatory response. Further studies found HDAC11 formed a complex with ERG and decreased the acetylation levels of ERG. More importantly, ERG knockdown augmented vascular endothelial cell pyroptosis in TNF-α-induced HUVECs. Taken together, our study suggests that HDAC11 might promote both NLRP3/caspase-1/GSDMD and caspase-3/GSDME pathways leading to pyroptosis via regulation of ERG acetylation in HUVECs. Modulation of HDAC11 may serve as a potential target for therapeutic strategies of AS.

Hydroxytyrosol Acetate Inhibits Vascular Endothelial Cell Pyroptosis via the HDAC11 Signaling Pathway in Atherosclerosis.

Hydroxytyrosol acetate (HT-AC), a natural polyphenolic compound in olive oil, exerts an anti-inflammatory effect in cardiovascular diseases (CVDs). Pyroptosis is a newly discovered form of programmed inflammatory cell death and is suggested to be involved in the atherosclerosis (AS) process. However, the effect of HT-AC on vascular endothelial cell pyroptosis remains unknown. Thus, we aimed to investigate the effect of HT-AC on vascular endothelial cell pyroptosis in AS and related signaling pathways. In vivo studies showed that HT-AC alleviated the formation of atherosclerotic lesions and inhibited pyroptosis in the aortic intima of ApoE-/- mice fed a high-fat diet (HFD) for 12 weeks. In vitro, we found that HT-AC treatment of human umbilical vein endothelial cells (HUVECs) alleviated tumor necrosis factor-alpha (TNF-α)-induced pyroptosis by decreasing the number of PI positive cells, decreasing the enhanced protein expressions of activated caspase-1 and gasdermin D (GSDMD), as well as by decreasing the release of pro-inflammatory interleukin (IL)-1ß and IL-6. Besides, HT-AC down-regulated HDAC11 expression in the aortic intima of HFD-fed ApoE-/- mice and TNF-α-stimulated HUVECs. To determine the underlying mechanism of action, molecular docking and drug affinity responsive target stability (DARTS) were utilized to identify whether HDAC11 protein is a target of HT-AC. The molecular docking result showed good compatibility between HT-AC and HDAC11. DARTS study's result showed that HDAC11 protein may be a target of HT-AC. Further study demonstrated that knockdown of HDAC11 augmented the inhibition of HT-AC on pyroptosis in TNF-α-stimulated HUVECs. These findings indicate that HT-AC might prevent vascular endothelial pyroptosis through down-regulation of HDAC11 related signaling pathway in AS.

Protective effect of SIRT6 on cholesterol crystal-induced endothelial dysfunction via regulating ACE2 expression.

Sirtuins are a family of highly conserved nicotinamide adenine dinucleotide (NAD+)-dependent enzymes. Among the sirtuins, SIRT1 and SIRT6 participate in the regulation of endothelial functions and play significant roles in the physiological and pathological processes of cardiovascular diseases (CVD). Recently, our study found that minute cholesterol crystals (CC) can be endocytosed by endothelial cells and further impair endothelial functions. Since previous studies have reported that angiotensin-converting enzyme (ACE2) involves Angiotensin (Ang) II-induced inflammation in endothelial cells, this study was designed to investigate the role of SIRT1 and SIRT6 in CC-induced variation of ACE2 expression and the related mechanism between SIRT6 and ACE2. We found that ACE2 is involved in CC-induced endothelial dysfunction, which inhibits decreases in nitric oxide (NO) level and endothelial nitric oxide synthase (eNOS) activity and increases in inflammatory factors and adhesion molecules. Besides, SIRT1 and SIRT6 regulated the protein expression of ACE2 in CC-stimulated human umbilical vein endothelial cells (HUVECs). Moreover, bioinformatics analysis from the Enrichr database indicated that activating transcription factor 2 (ATF2), is highly correlated with genes that significantly upregulated after infection with the SIRT6 adenovirus vector. In CC-induced HUVECs, ACE2 expression was up-regulated in cells transfected with ATF2 siRNA. However, further mechanism studies revealed that overexpression of SIRT6 decreases the accumulation of p-ATF2 in the nucleus, but did not affect p-ATF2 expression in the cytoplasm. Taken together, these data indicated that SIRT6 regulates ACE2 might via inhibiting the accumulation of nucleus p-ATF2 in CC-induced endothelial dysfunction.

A comparative study unraveling the effects of TNF-α stimulation on endothelial cells between 2D and 3D culture.

Endothelial cell (EC) dysfunction is an important predictor of and contributor to the pathobiology of cardiovascular diseases. However, most in vitro studies are performed using monolayer cultures of ECs on 2D tissue polystyrene plates (TCPs), which cannot reflect the physiological characteristics of cells in vivo. Here, we used 2D TCPs and a 3D culture model to investigate the effects of dimensionality and cardiovascular risk factors in regulating endothelial dysfunction. Cell morphology, oxidative stress, inflammatory cytokines and endothelial function were investigated in human umbilical vein endothelial cells (HUVECs) cultured in 2D/3D. The differentially expressed genes in 2D/3D-cultured HUVECs were analysed using Enrichr, Cytoscape and STRING services. Finally, we validated the proteins of interest and confirmed their relevance to TNF-α and the culture microenvironment. Compared with 2D TCPs, 3D culture increased TNF-α-stimulated oxidative stress and the inflammatory response and changed the mediators secreted by ECs. In addition, the functional characteristics, important pathways and key proteins were determined by bioinformatics analysis. Furthermore, we found that some key proteins, notably ACE, CD40, Sirt1 and Sirt6, represent a critical link between endothelial dysfunction and dimensionality, and these proteins were screened by bioinformatics analysis and verified by western blotting. Our observations provide insight into the interdependence between endothelial dysfunction and the complex microenvironment, which enhances our understanding of endothelial biology or provides a therapeutic strategy for cardiovascular-related diseases.

SIRT6 inhibits cholesterol crystal-induced vascular endothelial dysfunction via Nrf2 activation.

Sirtuin 6 (SIRT6), a nicotinamide adenine dinucleotide-dependent deacetylase, participates in various age-related disorders, such as dyslipidemia and cardiovascular diseases. Recent studies have revealed that minute cholesterol crystals (CCs), which are generated after excess free cholesterol accumulation, form not only in mature atherosclerotic plaques but also extremely early in atherosclerosis. Since endothelial dysfunction is an early feature of atherogenesis, this study was designed to investigate the role of SIRT6 in minute CC-induced endothelial dysfunction and the related mechanism. We found that minute CCs could be endocytosed by endothelial cells (ECs), which then decreased nitric oxide (NO) levels and endothelial nitric oxide synthase (eNOS) activity and expression, upregulated the expression of adhesion molecules and enhanced monocyte adhesion to ECs. In addition, minute CCs significantly suppressed SIRT6 expression in ECs. Moreover, the overexpression of SIRT6 could mitigate minute CC-induced endothelial dysfunction. In addition, the expression of Nuclear factor erythroid2-related factor2 (Nrf2) was suppressed after minute CC treatment, whereas SIRT6 overexpression reversed this decrease in Nrf2 expression. More importantly, Nrf2 activation also notably attenuated minute CC-induced endothelial dysfunction. In vivo experiments further indicated that endothelium-specific SIRT6 depletion impaired vascular endothelial function and suppressed Nrf2 expression in hyperlipidemic mice. Taken together, these results indicate that SIRT6 rescues minute CC-induced endothelial dysfunction partly via Nrf2 activation.

Sitagliptin inhibits vascular inflammation via the SIRT6-dependent signaling pathway.

Sitagliptin has recently been shown to inhibit inflammatory response in cardiovascular disease. Sirtuin6 (SIRT6), a NAD+-dependent class III histone deacetylase, participates in the regulation of cellular inflammation. We hypothesized that sitagliptin could attenuate vascular inflammation via modulation of SIRT6 pathway. It was found that sitagliptin decreased the expression of monocyte chemotactic protein-1 (MCP-1), interleukin (IL)-6 and IL-1ß, but up-regulated SIRT6 expression, both in mice and in TNF-α-stimulated endothelial cells. Moreover, knockdown of SIRT6 reversed the inhibitory effect of sitagliptin on MCP-1, IL-6 and IL-1ß expression. Further study revealed that sitagliptin also decreased the expression of MCP-1, IL-6 and IL-1ß partly through suppression of reactive oxygen species (ROS). In vivo, hypercholesterolemia in mice was induced by intraperitoneal administration of poloxamer 407 for 1 month. Hyperlipidemia-induced production of MCP-1, IL-6 and IL-1ß was significantly suppressed in the sitagliptin-supplemented animals, but the effect of was abolished by SIRT6 knockout in endothelium. These results indicate that sitagliptin protects against vascular inflammation via the SIRT6/ROS-dependent signaling pathway.

Carbonaceous nanofiber membrane functionalized by beta-cyclodextrins for molecular filtration.

In this paper, we report the fabrication of carbonaceous nanofiber (CNF) membranes functionalized by beta-cyclodextrins (CNF-ß-CD membrane) and their application for molecular filtration. The chemically synthesized carbonaceous nanofibers were first functionalized by ß-CD, and the free-standing CNF membrane can be prepared by a simple filtration process. The membrane shows a remarkable capability to function as an ideal molecular filter through complexation of phenolphthalein molecules with the cyclodextrin molecules grafted on the CNFs. As a typical dye pollutant, fuchsin acid can also be effectively removed from the solution through such a membrane. Engineering the surface of this carbonaceous nanofiber membrane may allow it to be used for other applications such as chiral separation and drug delivery.