Cold-inducible RNA-binding protein (CIRP/CIRBP) is recognized as an extracellular damage-associated molecular pattern. However, its relationship to endothelial redox-inflammatory injury in atherosclerosis remains poorly characterized. Here, we explored the associations among CIRP, endothelial dysfunction, and TLR4-/SIRT6-related changes. In high-fat diet (HFD)-fed ApoE-/- mice, circulating CIRP was elevated and positively correlated with atherosclerotic plaque burden. This increase coincided with systemic redox imbalance, impaired NO/eNOS activity, and vascular inflammation. In HUVECs, CIRP exposure reduced cell viability and impaired NO/eNOS function. These effects were accompanied by increased ROS accumulation and MDA content, together with reduced GSH-Px activity. CIRP also increased inflammatory cytokine production, NF-κB p65 phosphorylation, and THP-1 adhesion. At the molecular level, CIRP reduced SIRT6 expression. Overexpression of SIRT6 attenuated CIRP-induced endothelial injury, oxidative stress, and NO/eNOS dysfunction. CIRP also increased TLR4 expression. Accordingly, pharmacological TLR4 inhibition with TAK-242 attenuated CIRP-associated endothelial injury and partially restored SIRT6 expression and protein stability. Conversely, SIRT6 silencing weakened the protective effects of TAK-242. In addition, CIRP exposure was accompanied by increased overall eNOS acetylation. This increase was attenuated by TAK-242 and further enhanced by SIRT6 silencing. Exploratory cross-context transcriptomic analysis identified overlapping inflammatory and oxidative stress-related signatures. Representative antioxidant-related changes were further examined in CIRP-treated HUVECs through assessment of GPX4 and CAT mRNA expression and CAT activity. Finally, in a preliminary clinical cohort, serum CIRP levels were higher in patients with coronary heart disease and were positively associated with Gensini score, including selected exploratory multivariable models. Overall, these findings provide preliminary evidence that CIRP is associated with redox-inflammatory endothelial injury. The results are also consistent with the possible involvement of TLR4-/SIRT6-related signaling. These observations should be interpreted cautiously because of the relatively high CIRP concentration used in vitro, the cross-context transcriptomic comparison, and the small clinical cohort.

BACKGROUND & AIMS:Metabolic dysfunction-associated steatotic liver disease (MASLD) is a major global health concern, with obesity serving as a primary risk factor. Although Mediator subunit 1 (MED1) plays an important role in lipid metabolism, its specific contribution to obesity-related hepatic steatosis remains unclear. This study aims to elucidate the involvement of MED1 in the pathogenesis of MASLD during obesity. APPROACH & RESULTS:Herein, we found that MED1 expression was upregulated in fatty livers from obese patients with MASLD, a primate MASLD model, genetically obese (ob/ob) mice, and in palmitate-treated HepG2 cells. Hepatocyte-specific knockout of MED1 on an ob/ob background under both chow and high-fat diet feeding ameliorated hepatic steatosis, glucose intolerance, obesity and inflammation of visceral white adipose tissue. Mechanistically, MED1 regulates hepatic lipid metabolism primarily through direct interaction with SREBP1, thereby mediating the expression of key SREBP1 target genes, including ACC, FASN and SCD1. Importantly, therapeutic delivery of AAV8-shMED1 attenuated MASLD progression in ob/ob mice. CONCLUSIONS:These findings establish MED1 as a critical activator of SREBP1-driven lipogenesis and identify hepatic MED1 inhibition as a promising therapeutic strategy for MASLD.

BACKGROUND:Currently, no drug treatment is available for abdominal aortic aneurysms (AAAs). Berberine, an alkaloid extracted from the traditional Chinese herbs Coptis chinensis and Phellodendron amurense, has several beneficial biological effects, including anti-inflammatory properties. PURPOSE:The aim of this study was to explore whether berberine affects the pathogenesis of AAAs. METHODS:An intraluminal porcine pancreatic elastase (PPE) infusion-induced mouse model of AAAs and AI-based analysis were used to identify the efficacy of berberine in the inhibition of experimental AAAs. RESULTS:We found that berberine treatment significantly inhibited PPE-induced aortic dilation. Histopathologic analysis revealed that berberine treatment reduced PPE-induced inflammatory cell infiltration in the aortic wall, promoted vascular smooth muscle cell (VSMC) survival, and protected against medial elastin degradation. Moreover, berberine treatment also downregulated matrix metalloproteinase 2 (MMP-2) and MMP-9 expression in the aortic wall and reduced abnormal mural angiogenesis. Through the use of public databases and bioinformatics, machine learning, and molecular docking techniques, RUNX2, VCAM-1, and CCL2 were identified as hub genes through which berberine inhibits AAAs. To confirm the above results, we performed tissue-specific knockout of Runx2 in VSMCs and found that Runx2 deficiency attenuated PPE-induced AAAs by increasing medial smooth muscle cell depletion, elastin degradation, and aortic inflammation. CONCLUSION:In conclusion, berberine has an anti-aneurysmal effect, which may be related to the protection of VSMCs and elastin related to positive vascular remodeling and the inhibition of abnormal aortic inflammation. This study is the first to demonstrate the anti-aneurysmal effect of berberine, which may provide a new potential option for treating AAAs.

Histone deacetylase 11 (HDAC11) is the only member of the class IV HDAC family and is involved in cardiovascular diseases (CVDs). Stress granule (SG) is non-membranous cytoplasmic foci induced by various stress conditions, and also has emerged as a key player for CVDs. However, the regulatory role of HDAC11 in SG formation and underlying mechanism during atherosclerosis remain elusive. Therefore, we aimed to investigate the effect of HDAC11 on SG in ApoE-/- mice fed with a HFD and HUVECs induced by H2O2. Firstly, we found that the expression levels of SG core proteins G3BP1/2 and HDAC11 were increased in the aorta of ApoE-/- mice fed with a HFD for 12w via analyses of Western blotting, Real-time PCR and immunofluorescence staining. In addition, endothelial-to-mesenchymal transition (EndMT) was occurred in the aorta of ApoE-/- mice. Then, in vitro experiments demonstrated that treatment of HUVECs with H2O2 resulted in SG formation, HDAC11 upregulation, and EndMT occurrence. Furthermore, knockdown of HDAC11 by siRNA significantly attenuated SG formation and EndMT activation in HUVECs induced by H2O2. Silencing of HDAC11 suppressed H2O2-induced EndMT activation in HUVECs, which may be attributed to increased acetylation of G3BP1/2 and the consequent impairment of SG formation. Further studies found that suppression of SG formation not only facilitated the expression of endothelial markers, but also decreased the levels of mesenchymal cell markers. Taken together, these findings identified that HDAC11 may regulate SG formation to promote EndMT in atherosclerosis, targeting SG could represent a novel therapeutic strategy for addressing the underlying mechanisms of atherosclerosis.

Histone deacetylase 3 (HDAC3) is an epigenetic modifying enzyme closely linked to the development of atherosclerosis. Endothelial inflammation is a critical factor in atherosclerosis. However, the role of HDAC3 in mediating epigenetic modifications and regulating endothelial inflammation in atherosclerosis remains unclear. This study aims to investigate the impact of HDAC3 on endothelial inflammation and its contribution to atherosclerosis. Firstly, single-cell transcriptomic analysis identified elevated expression of HDAC3 and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) in inflammatory endothelial cells of atherosclerotic plaques in symptomatic patients. Endothelial-specific knockout HDAC3 in an apolipoprotein E knockout (ApoE−/−) mice decreased atherosclerotic lesion by reducing lipid deposition and endothelial NLRP3 inflammasome activation compared with control mice. Consistently, experiments using HDAC3 inhibitor and overexpression in human umbilical vein endothelial cells (HUVECs) demonstrated that HDAC3 enhanced the transcriptional upregulation of NLRP3 inflammasome by promoting nuclear factor kappa-B pathway, thereby contributing to the activation of the NLRP3 inflammasome and cellular injury. Further studies revealed that HDAC3 reduced specificity protein 1 (SP1) Lys-703 acetylation, thereby enhancing SP1 binding to the NLRP3 promoter and promoting NLRP3 transcription. Additionally, pharmacological inhibition of HDAC3 effectively ameliorated atherosclerosis by reducing endothelial inflammation and increasing SP1 acetylation in ApoE−/− mice. Thus, these findings demonstrate a crucial role of HDAC3 in endothelial inflammation and shed light on potential therapeutic strategy for atherosclerosis via inhibition of HDAC3.

An abnormal inflammatory response is one of the main pathogenic mechanisms of abdominal aortic aneurysms (AAAs), and tranilast, an antiallergic drug, has anti-inflammatory properties. The effect and mechanism of action of tranilast on AAAs remain incompletely defined. To evaluate the preventive and therapeutic effects on experimental AAAs induced by intra-aortic elastase infusion in mice, tranilast was administered either before or after elastase infusion and continued until the experimental endpoint. Bioinformatics analysis and corresponding validation experiments were used to explore the possible mechanisms by which tranilast affects AAA progression. Compared with vehicle treatment, both tranilast pre-treatment and post-treatment therapies markedly inhibited aneurysmal aortic expansion. Treatment with tranilast attenuated the degradation of aneurysmal medial elastin and the depletion of smooth muscle cells. Aortic leukocyte accumulation was significantly lower in aneurysmal aortas from tranilast-treated mice than in those from vehicle-treated mice. Mural abnormal angiogenesis and aortic matrix metalloproteinase (MMP) 2 and 9 expression levels were also reduced after tranilast treatment. Bioinformatics analysis revealed that nucleotide-binding oligomerization domain-like receptor family protein 3 (NLRP3) may be a hub target through which tranilast affects AAAs. NLRP3 expression levels were lower in the aneurysmal aortas of tranilast-treated mice than in those of vehicle-treated elastase-infused mice. Both Nlrp3 deficiency and treatment with the NLRP3 inhibitor MCC950 attenuated experimental AAAs. However, cotreatment with tranilast had no additive or synergistic influence on AAA suppression. Additionally, tranilast treatment reduced caspase 1 cleavage by the NLRP3 inflammasome and consequently interleukin-1β secretion in peritoneal macrophages in vitro. These findings indicate that the protective effect of tranilast on AAA may be partially mediated by the inhibition of the NLRP3 inflammasome pathway and may represent a potential drug for the treatment of AAA in the clinic.

Background and purpose: Catalpol (CAT) has diverse pharmacological functions, including cellular homeostasis maintenance and anti-inflammatory effects. Sirtuin 5 (SIRT5) plays a considerable role in regulating cellular homeostasis in cardiac diseases. Our research explores the therapeutic potential of CAT against myocardial injury and its underlying mechanism. Methods: The H9c2 cells were pretreated with different CAT concentrations for 24 h, or CAT for 24 h followed by CoCl2 stimulation. Cell viability was determined with MTT assay. Biochemical assays, western blotting, and quantitative real-time PCR (qRT-PCR), combined with bioinformatic analysis, were used to examine the impact of CAT on CoCl2-induced myocardial injury in H9c2 cells and further explore its molecular mechanisms. Results: CAT ameliorated levels of myocardial enzymes, increased nicotinamide adenine dinucleotide (NAD+/NADH) ratio and adenosine triphosphate (ATP), while inhibited lactic acid (LD), tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta, and IL-6 in CoCl2-induced H9c2 cells. Mechanistically, SIRT5 knockdown inhibited Lin28a expression and negated the effects of CAT on ATP level, LD content, and the expression of inflammatory factors in cells. CAT likely exerted its protective effects on myocardial function through the SIRT5-mediated signaling pathway. Conclusions: CAT regulates energy metabolism and inflammation via the SIRT5-mediated signaling pathway, exerting a protective effect in myocardial injury.

Background Diabetes mellitus (DM) and its complications seriously threaten human life and health. Rhaponticum carthamoides (Willd.) Iljin (RC) is widely used to treat cardiovascular diseases. Previous studies reported that RC reduces blood glucose levels in rats with type 1 DM. However, the effects of RC on type 2 diabetes and vascular complications, as well as its related active components and underlying mechanisms, remain unclear. Purpose This study aimed to investigate the effects of RC on endothelial dysfunction and the inflammatory response in type 2 DM mice and to explore its underlying mechanism and active ingredients. Study Design/Methods Male C57BL/6J mice were used to establish a type 2 DM mouse model. After 12 weeks of oral administration of RC extract (60, 120, and 240 mg/kg) to mice, blood glucose and lipid levels were assessed. The morphological structures of the liver and kidney tissues were observed using hematoxylin and eosin (HE) staining, and their functions were evaluated by detecting relevant biochemical indicators in the serum. Then, aorta morphology was observed via HE staining. In addition, serum levels of markers of endothelial function and inflammatory factors were detected, and the expression of inflammatory factors and the phosphorylation levels of key proteins in the aorta were examined. Furthermore, prediction and enrichment analyses of potential targets of RC acting on diabetic vascular lesions were performed on the basis of pharmacophore matching using various databases. Then, the expression, localization and phosphorylation levels of potential targets in the aortas of DM mice treated with RC were assessed using Western blotting, immunofluorescence, and RT‒PCR. Finally, the active components of RC were identified through virtual screening, and their ability to improve endothelial cell dysfunction was verified. Results RC reduced blood glucose levels and serum lipid levels of total triglyceride (TG), total cholesterol (TC), and low density lipoprotein cholesterol (LDL-c), increased high density lipoprotein cholesterol (HDL-c) levels, and improved liver and kidney function in type 2 DM mice. RC decreased endothelial cell shedding in the aortas of type 2 DM mice, increased serum nitric oxide (NO) and nitric oxide synthase (NOS) levels, and reduced soluble cluster of differentiation 40 ligand (sCD40L), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β) levels. Further findings indicated that RC reduced the expression of aortic inflammatory factors, namely, CD40, CD40L, IL-1β, and interleukin-6 (IL-6), and increased endothelial NOS (eNOS) phosphorylation levels. Sirtuin 6 (SIRT6), protein kinase B (AKT), and eNOS were predicted to be key node targets of RC acting on DM vascular lesions, and it was confirmed that RC increased SIRT6 expression and AKT phosphorylation levels in aortic endothelial cells. 20-Hydroxyecdysone (20E), daucosterol (Dau), euscaphic acid (Eus), and syringin (Syr) were identified as active components of RC. These components protect against TNF-α-induced human umbilical vein endothelial cell (HUVEC) damage and decrease the release of lactate dehydrogenase (LDH) and IL-1β and increased the release of NO in TNF-α-induced HUVECs in a dose-dependent manner. Conclusion RC reduced blood glucose and lipid levels in mice with type 2 DM and protected liver and kidney function. RC promotes SIRT6 expression in endothelial cells; upregulates the NO/NOS system by increasing AKT/eNOS phosphorylation levels to regulate vascular tone factors; and reduces the levels of inflammatory factors such as CD40, TNF-α, and IL-1β to inhibit endothelial inflammatory responses. Based on these mechanisms, RC improves endothelial dysfunction.

Intimal hyperplasia (IH) is a common pathological feature of vascular proliferative diseases, such as atherosclerosis and restenosis after angioplasty. Urotensin II (UII) and its receptor (UTR) are widely expressed in cardiovascular tissues. However, it remains unclear whether the UII/UTR system is involved in IH. Right unilateral common carotid artery ligation was performed and maintained for 21 days to induce IH in UTR knockout (UTR-/-) and wild-type (WT) mice. Histological analysis revealed that compared with WT mice, UTR-deficient mice exhibited a decreased neointimal area, angiostenosis and intima-media ratio. Immunostaining revealed fewer smooth muscle cells (SMCs), endothelial cells and macrophages in the lesions of UTR-/- mice than in those of WT mice. Protein interaction analysis suggested that the UTR may affect cell proliferation by regulating YAP and its downstream target genes. In vitro experiments revealed that UII can promote the proliferation and migration of SMCs, and western blotting also revealed that UII increased the protein expression of RhoA, CTGF, Cyclin D1 and PCNA and downregulated p-YAP protein expression, while these effects could be partly reversed by urantide. To evaluate the translational value of UTRs in IH management, WT mice were also treated with two doses of urantide, a UTR antagonist, to confirm the benefit of UTR blockade in IH progression. A high dose of urantide (600 μg/kg/day), rather than a low dose (60 μg/kg/day), successfully improved ligation-induced IH compared with that in mice receiving vehicle. The results of the present study suggested that the UII/UTR system may regulate IH partly through the RhoA-YAP signaling pathway.

Histone deacetylases (HDACs) play critical roles in cardiovascular diseases (CVDs). In addition, reactive oxygen species (ROS) produced by NADPH oxidases (NOXs) exert damaging effects due to oxidative stress on heart and blood vessels. Although NOX-dependent ROS production is implicated in pathogenesis, the relationship between HDACs and NOXs in CVDs remains to be elucidated. Here, we present an overview of the regulatory effects and interconnected signaling pathways of HDACs and NOXs in CVDs. Improved insights into these relationships will facilitate the discovery of novel therapeutic agents that target HDACs, oxidase stress pathways, and the interactions between these systems which may be highly effective in the prevention and treatment of cardiovascular disorders.

INTRODUCTION:Kaempferol (KAE) is a flavonoid found in various plants. Recent studies showed that high dietary intake of KAE was associated with a lower risk of myocardial infarction; however, the cardioprotective mechanism of KAE remains unknown. OBJECTIVES:To determine the effect of KAE on cardiac injury in isoproterenol (ISO)-induced rats and cobalt chloride (CoCl2)-treated cardiomyocytes, and the underlying mechanisms. METHODS:Male rats were pretreated with different doses of KAE for 14 days, and then injected with ISO to induce myocardial ischemia injury. We also established a model of myocardial cell injury using rat H9c2 cardiomyocytes stimulated with CoCl2. RESULTS:We found that KAE pretreatment significantly alleviated myocardial injury and improved cardiac function in ISO-injected rats. In addition, KAE reduced oxidative stress in rats with myocardial ischemia by decreasing malondialdehyde concentration and increasing superoxide dismutase activity, and protection of the myocardial mitochondrial structure. KAE also attenuated CoCl2-induced injuryof H9c2 cardiomyocytes via suppression ofoxidative stress. With regard to the mechanism, we found that KAE down-regulated HDAC3 expression and up-regulated Nrf2 expression in ISO-induced rats and CoCl2-stimulated cardiomyocytes. Incubation of cardiomyocytes with HDAC3-selective inhibitor RGFP966 augmented the protective effect of KAE and reduced oxidative stress. By contrast, HDAC3 overexpression by adenovirus attenuated the effect of KAE on oxidative stress compared with KAE treatment group. HDAC3 also regulated Nrf2 expression in the cardiomyocytes with RGFP966 or an adenovirus overexpressing HDAC3; but Nrf2 inhibition reduced the effect of KAE on ROS generation in CoCl2-induced cardiomyocytes. Immunoprecipitation assay showed that HDAC3 interacted with Nrf2 in cardiomyocytes. Further studies found that KAE increased the acetylation level of Nrf2, while HDAC3 overexpression decreased the acetylation of Nrf2 compared with KAE treatment group. CONCLUSION:Our data show that KAE ameliorates cardiac injury by reducing oxidative stress via the HDAC3-mediated Nrf2 signaling pathway in cardiomyocytes.

BACKGROUND AND AIMS:Phenotypic switching of vascular smooth muscle cells (VSMCs) plays an essential role in the development of atherosclerosis. Protein inhibitor of activated STAT (Pias) regulates VSMCs phenotype via acting as sumo E3 ligase to promote protein sumoylation. Our previous study indicated that Pias3 expression decreased in atherosclerotic lesions. Therefore, this study aimed to explore the role of Pias3 on VSMCs phenotype switching during atherosclerosis.METHODS:ApoE-/- and ApoE-/-Pias3-/- double-deficient mice were fed with high-fat/high-cholesterol diet to induce atherosclerosis. Aorta tissues and primary VSMCs were collected to assess plaque formation and VSMCs phenotype. In vitro, Pias3 was overexpressed in A7r5, a VSMCs cell line, by transfection with Pias3 plasmid. Real-time quantitative PCR, immunoblotting, immunoprecipitation, were used to analyze the effect of Pias3 on VSMCs phenotypic switching.RESULTS:Pias3 deficiency significantly exacerbated atherosclerotic plaque formation and promoted VSMCs phenotypic switching to a synthetic state within lesion. In vitro, overexpressing Pias3 in VSMCs increased the expression of contractile markers (myosin heavy chain 11, calponin 1), while it decreased the level of synthetic marker (vimentin). Additionally, Pias3 overexpression blocked PDGF-BB-induced VSMCs proliferation and migration. Immunoprecipitation and mass spectrometry results showed that Pias3 enhanced sumoylation and ubiquitination of vimentin, and shortened its half-life. Moreover, the ubiquitination level of vimentin was impaired by 2-D08, a sumoylation inhibitor. This suggests that Pias3 might accelerate the ubiquitination-degradation of vimentin by promoting its sumoylation.CONCLUSIONS:These results indicate that Pias3 might ameliorate atherosclerosis progression by suppressing VSMCs phenotypic switching and reducing vimentin protein stability.

In this study, we aimed to analyze the proteomics of the liver in rabbits on a high cholesterol diet (HCD). We randomly divided New Zealand white rabbits into the normal diet group and the HCD group. We established the atherosclerosis model and measured plasma cholesterol and triglycerides. The model was successfully established using ultrasound examination and histopathological staining of the intima of aorta and liver of the two groups of rabbits. The differential proteins in the rabbit liver were analyzed using Tandem Mass Tags proteomic analysis technology. Finally, we used western blot to verify the reliability of proteomics. The results showed that compared with the control group, the serum lipid levels of rats in the HCD group was significantly increased, and the pathological sections showed the formation of atherosclerotic plaques in the aorta, inflammation, and adipose lesions in the liver. Proteomic analysis of the liver revealed 149 differences in HCD-expressed protein, which is mainly involved in inflammation and regulation of lipid and sugar metabolism. In addition, we verified differentially expressed liver proteins in the HCD group using western blot. We found that HCD caused lipid accumulation, abnormal glucose metabolism, and inflammatory response in the liver.

Background Endothelial-to-mesenchymal transition (EndMT) is the process by which endothelial cells lose their specific markers and acquire mesenchymal or myofibroblastic phenotypes. Studies have demonstrated the importance of endothelial-derived vascular smooth muscle cells (VSMCs) through EndMT in neointimal hyperplasia. Histone deacetylases (HDACs) are epigenetic modification enzymes involved in the epigenetic control of important cellular functions. Recent studies found that HDAC3, a class I HDAC, causes posttranslational modifications, including deacetylation and decrotonylation. However, the effect of HDAC3 on EndMT in neointimal hyperplasia via posttranslational modifications remains to be seen. Therefore, we investigated the effects of HDAC3 on EndMT in carotid artery-ligated mice and human umbilical vein endothelial cells (HUVECs) and the underlying posttranslational modifications. Methods HUVECs were treated with transforming growth factor (TGF)-β1 or the inflammatory cytokine tumor necrosis factor (TNF)-α at different concentrations and durations. In HUVECs, HDAC3 expression, the expression of endothelial and mesenchymal markers, and posttranslational modifications were analyzed with Western blotting, quantitative real-time polymerase chain reaction (PCR), and immunofluorescence. C57BL/6 mice underwent left carotid artery ligation. Mice were treated with the HDAC3-selective inhibitor RGFP966 (10 mg/kg, i.p.) from 1 day before to 14 days after ligation. Then, the sections of the carotid arteries were examined histologically using hematoxylin and eosin (HE) and immunofluorescence staining. The carotid arteries from other mice were examined for the expression of EndMT markers and inflammatory cytokines. Furthermore, the acetylation and crotonylation of carotid arteries were immunostained in mice. Results In HUVECs, TGF-β1 and TNF-α induced EndMT by decreasing CD31 expression and increasing α-smooth muscle actin expression. TGF-β1 and TNF-α also upregulated HDAC3 expression in HUVECs. The in vivo study in mice indicated that RGFP966 significantly alleviated neointimal hyperplasia of the carotid artery compared with vehicle treatment. Furthermore, RGFP966 suppressed EndMT and the inflammatory response in carotid artery-ligated mice. Further investigation revealed that HDAC3 regulated EndMT by posttranslational modifications of deacetylation and decrotonylation. Conclusions These results suggest that HDAC3 regulates EndMT in neointimal hyperplasia through posttranslational modifications.

BackgroundC-reactive protein (CRP) levels are elevated in patients with abdominal aortic aneurysms (AAA). However, it has not been investigated whether CRP contributes to AAA pathogenesis.MethodsCRP deficient and wild type (WT) male mice were subjected to AAA induction via transient intra-aortic infusion of porcine pancreatic elastase. AAAs were monitored by in situ measurements of maximal infrarenal aortic external diameters immediately prior to and 14 days following elastase infusion. Key AAA pathologies were assessed by histochemical and immunohistochemical staining procedures. The influence of CRP deficiency on macrophage activation was evaluated in peritoneal macrophages in vitro.ResultsCRP protein levels were higher in aneurysmal than that in non-aneurysmal aortas. Aneurysmal aortic dilation was markedly suppressed in CRP deficient (aortic diameter: 1.08 ± 0.11 mm) as compared to WT (1.21 ± 0.08 mm) mice on day 14 after elastase infusion. More medial elastin was retained in CRP deficient than in WT elastase-infused mice. Macrophage accumulation was significantly less in aneurysmal aorta from CRP deficient than that from WT mice. Matrix metalloproteinase 2 expression was also attenuated in CRP deficient as compared to WT aneurysmal aortas. CRP deficiency had no recognizable influence on medial smooth muscle loss, lymphocyte accumulation, aneurysmal angiogenesis, and matrix metalloproteinase 9 expression. In in vitro assays, mRNA levels for tumor necrosis factor α and cyclooxygenase 2 were reduced in lipopolysaccharide activated peritoneal macrophages from CRP deficient as compared to wild type mice.ConclusionCRP deficiency suppressed experimental AAAs by attenuating aneurysmal elastin destruction, macrophage accumulation and matrix metalloproteinase 2 expression.

Accumulated evidence shows that elevated urotensin II (UII) levels are associated with cardiovascular diseases. However, the role of UII in the initiation, progression, and regression of atherosclerosis remains to be verified. Different stages of atherosclerosis were induced in rabbits by a 0.3% high cholesterol diet (HCD) feeding, and either UII (5.4 μg/kg/h) or saline was chronically infused via osmotic mini-pumps. UII promoted atherosclerotic fatty streak formation in ovariectomized female rabbits (34% increase in gross lesion and 93% increase in microscopic lesion), and in male rabbits (39% increase in gross lesion). UII infusion significantly increased the plaque size of the carotid and subclavian arteries (69% increase over the control). In addition, UII infusion significantly enhanced the development of coronary lesions by increasing plaque size and lumen stenosis. Histopathological analysis revealed that aortic lesions in the UII group were characterized by increasing lesional macrophages, lipid deposition, and intra-plaque neovessel formation. UII infusion also significantly delayed the regression of atherosclerosis in rabbits by increasing the intra-plaque macrophage ratio. Furthermore, UII treatment led to a significant increase in NOX2 and HIF-1α/VEGF-A expression accompanied by increased reactive oxygen species levels in cultured macrophages. Tubule formation assays showed that UII exerted a pro-angiogenic effect in cultured endothelial cell lines and this effect was partly inhibited by urantide, a UII receptor antagonist. These findings suggest that UII can accelerate aortic and coronary plaque formation and enhance aortic plaque vulnerability, but delay the regression of atherosclerosis. The role of UII on angiogenesis in the lesion may be involved in complex plaque development.

目的 研究异甘草素对血管内皮细胞炎症反应的保护作用,以及异甘草素是否通过组蛋白去乙酰化酶 3(his-tone deacetylase 3,HDAC3)调控血管内皮细胞的炎症反应.方法 以人脐静脉内皮细胞(HUVECs)为研究对象,分别给予脂多糖(LPS)刺激、不同浓度异甘草素联合 LPS处理细胞、HDAC3 特异性抑制剂和异甘草素联合 LPS处理细胞,Real-time PCR和 Western blotting检测细胞炎症因子和 HDAC3 的 mRNA和蛋白表达.雄性 C57 BL/6J 小鼠随机分为溶剂对照组和异甘草素组,颈动脉结扎手术建立小鼠急性血管炎症模型,Real-time PCR检测小鼠颈动脉炎症因子和 HDAC3 的mRNA表达.利用分子对接模型,从分子结构上揭示异甘草素和 HDAC3 之间的结合程度.结果 与溶剂对照组相比,异甘草素组中 LPS刺激所升高的血管内皮细胞炎症因子 NLRP3、IL-1β、IL-18、MCP-1 和 ICAM-1的 mRNA表达降低,细胞 HDAC3 的 mRNA和蛋白表达降低.此外,颈动脉结扎的手术组小鼠中,异甘草素组的血管炎症因子 NLRP3、IL-1β和 HDAC3 的 mRNA表达降低.分子对接结果显示异甘草素与 HDAC3 之间具有结构上的契合性,而且 HDAC3 特异性抑制剂 RGFP966 能够进一步促进异甘草素对血管内皮细胞炎症因子表达的抑制作用.结论 异甘草素可能通过 HDAC3 抑制血管内皮细胞的炎症反应.

The cultivation of innovation ability of medical students is an important part of reform and innovation,and a major way to improve the quality of medical talents in medical universities.Medical Laboratory Animal Science specializes in medical laboratory animals and animal experiments.It is an important basic course for medical students in colleges and universities.To better cultivate medical students'innovation ability in the teaching process of Medical Laboratory Animal Science,we introduce the scientific research case teaching method into the teaching of theoretical course,which closely combines the theoretical teaching contents with the teachers'scientific research.In addition,we reform the teaching contents according to the progress of subject,and integrate the latest scientific research into our teaching to cultivate students'innovative thinking ability.To promote the cultivation of students'scientific research and innovation ability,we strengthen the training of students'basic operation skills in animal experiments,and combine the experimental teaching with the innovation fund project of college students during the experimental course.To give full play to the advantages of Medical Laboratory Animal Science teaching in cultivating students'innovative ability and meet the requirements of innovative talent cultivation,we improve the course evaluation system and add the as-signment of animal experiment design in the course assessment.Therefore,the medical students are transformed from passive learning to active thinking and the initiative of learning is enhanced.Furthermore,we carry out special animal welfare and ethics education and strengthen the animal welfare concept throughout the whole teaching process to improve the humanistic quality of medical students.Through carrying out the innovative education for medical students and promoting the cultivation of innovative talents in the teaching process of Medical Laboratory Animal Science,we lay a solid foundation for cultivating medical innovative talents to meet the requirements of the new era.

20-Hydroxyecdysone (20E) is known to have numerous pharmacological activities and can be used to treat diabetes and cardiovascular diseases. However, the protective effects of 20E against endothelial dysfunction and its targets remain unclear. In the present study, we revealed that 20E treatment could modulate the release of the endothelium-derived vasomotor factors NO, PGI2 and ET-1 and suppress the expression of ACE in TNF-α-induced 3D-cultured HUVECs. In addition, 20E suppressed the expression of CD40 and promoted the expression of SIRT6 in TNF-α-induced 3D-cultured HUVECs. The cellular thermal shift assay (CETSA), drug affinity responsive target stability (DARTS) and molecular docking results demonstrated that 20E binding increased SIRT6 stability, indicating that 20E directly bound to SIRT6 in HUVECs. Further investigation of the underlying mechanism showed that 20E could upregulate SIRT6 levels and that SIRT6 knockdown abolished the regulatory effect of 20E on CD40 in TNF-α-induced HUVECs, while SIRT6 overexpression further improved the effect of 20E. Moreover, we found that 20E could reduce the acetylation of NF-κB p65 (K310) through SIRT6, but the catalytic inactive mutant SIRT6 (H133Y) did not promote the deacetylation of NF-κB p65, suggesting that the inhibitory effect of 20E on NF-κB p65 was dependent on SIRT6 deacetylase activity. Additionally, our results indicated that 20E inhibited NF-κB via SIRT6, and the expression of CD40 was increased in HUVECs treated with SIRT6 siRNA and NF-κB inhibitor. In conclusion, the present study demonstrates that 20E exerts its effect through SIRT6-mediated deacetylation of NF-κB p65 (K310) to inhibit CD40 expression in ECs, and 20E may have therapeutic potential for the treatment of cardiovascular diseases.

Background Rabbits are well-domesticated animals. As a crucial economic animal, rabbit has been successfully bred into wool-use, meat-use and fur-use breeds. Hair length is one of the most economically important traits affecting profitability in wool rabbits. In this study, to identify selection signatures with the long-hair trait, whole-genomic resequencing of long-haired rabbits (Angora rabbits) and short-haired rabbits (Rex and New Zealand rabbits) was performed. Results By genome-wide selective sweeping analysis based on population comparison, we identified a total of 5.85 Mb regions (containing 174 candidate genes) with strong selection signals. Six of these genes ( Dusp1 , Ihh , Fam134a , Map3k1 , Spata16 , and Fgf5 ) were enriched in the MAPK signalling and Hedgehog signalling pathways, both of which are closely associated with hair growth regulation. Among these genes, Fgf5 encodes the FGF5 protein, which is a well-established regulator of hair growth. There was a nonsynonymous nucleotide substitution (T19234C) in the Fgf5 gene. At this locus, the C allele was present in all of the tested Angora rabbits, while the T allele was dominant in New Zealand and Rex rabbits. We further confirmed that the C allele was conserved in Angora rabbits by screening an additional 135 rabbits. Moreover, the results of functional predictions and co-immunoprecipitation revealed that the T19234C mutation impaired the binding capacity of FGF5 to its receptor FGFR1. Conclusions We discovered that the homozygous missense mutation T19234C within Fgf5 might contribute to the long-hair trait of Angora rabbits by reducing its receptor binding capacity. This finding will provide new insights into the genetic basis underlying the genetic improvement of Angora rabbits and benefit the improvement of rabbit breeding in the future.