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The evolution of endovascular therapies, particularly in the field of intracranial aneurysm treatment, has been truly remarkable and is characterized by the development of various stents. However, ischemic complications related to thrombosis or downstream emboli pose a challenge for the broader clinical application of such stents. Despite advancements in surface modification technologies, an ideal coating that fulfills all the desired requirements, including anti-thrombogenicity and swift endothelialization, has not been available. To address these issues, we investigated a new coating comprising 3-aminopropyltriethoxysilane (APTES) with both anti-thrombogenic and cell-adhesion properties. We assessed the anti-thrombogenic property of the coating using an in vitro blood loop model by evaluating the platelet count and the level of the thrombin-antithrombin (TAT) complex, and investigating thrombus formation on the surface using scanning electron microscopy (SEM). We then assessed endothelial cell adhesion on the metal surfaces. In vitro blood tests revealed that, compared to a bare stent, the coating significantly inhibited platelet reduction and thrombus formation; more human serum albumin spontaneously adhered to the coated surface to block thrombogenic activation in the blood. Cell adhesion tests also indicated a significant increase in the number of cells adhering to the APTES-coated surfaces compared to the numbers adhering to either the bare stent or the stent coated with an anti-fouling phospholipid polymer. Finally, we performed an in vivo safety test by implanting coated stents into the internal thoracic arteries and ascending pharyngeal arteries of minipigs, and subsequently assessing the health status and vessel patency of the arteries by angiography over the course of 1 week. We found that there were no adverse effects on the pigs and the vascular lumens of their vessels were well maintained in the group with APTES-coated stents. Therefore, our new coating exhibited both high anti-thrombogenicity and cell-adhesion properties, which fulfill the requirements of an implantable stent.

Objectives: Deep-venous thrombosis (DVT) refers to abnormal blood clotting in the deep vein cavity, and post-thrombotic syndrome (PTS) is the most frequent complication. The study explored the impact of microRNA 181b-5p on DVT progression based on human umbilical vein endothelial cells (HUVECs). Methods: Levels of miR-181b-5p were examined in 150 cases with acute lower extremity DVT. ROC curve and K-M plot were drawn for clinical value assessment. The role of miR-181b-5p in HUVECs viability, migration, apoptosis, inflammatory response and adhesion factors' release was investigated. Target gene of miR-181b-5p was predicted, and its role in cell function was explored. Results: Low-expressed miR-181b-5p showed favorable diagnostic performance in differentiating DVT with the AUC of 0.948. Patients with low miR-181b-5p had a high incidence of PTS. miR-181b-5p overexpression promoted HUVECs' viability and migration, while inhibiting cell apoptosis and release of inflammatory and adhesion cytokines. As the target gene of miR-181b-5p, HEY2 overexpression reversed the role of miR-181b-5p in HUVECs. Conclusion: MiR-181b-5p serves as a potential biomarker for DVT diagnosis and PTS development. Overexpression of this miRNA targeted HEY2 to alleviate endothelial cell damage. [ABSTRACT FROM AUTHOR]

Angiogenesis is associated with angiogenic therapy and wound healing processes. It is important for anesthesiologists to understand the effects of perioperative and long-term use of anesthetics on angiogenesis. This study aimed to determine the effects of ketamine on in vitro angiogenesis: the proliferation of human umbilical vein endothelial cells (HUVEC) and normal human diploid fibroblasts (NHDF), HUVEC migration, and in vitro capillary tube formation in cocultured HUVEC and NHDF. The effects of ketamine at concentrations of 1, 10, and 50 µM on the proliferation of HUVEC and NHDF for 48 hours were determined by using a water-soluble tetrazolium salt reagent. Quantitation of migration for 22 hours was achieved by measuring the fluorescence of migrating HUVEC exposed to ketamine using an angiogenesis system. The effects of ketamine on capillary tube formation with or without vascular endothelial growth factor (VEGF) were investigated in cocultured HUVEC and NHDF incubated for 3 and 10 days. Ketamine did not show any enhancing or suppressive effects on the in vitro proliferation of HUVEC and NHDF, HUVEC migration, or capillary tube formation in cocultured HUVEC and NHDF for either 3 or 10 days in the presence or absence of VEGF. Ketamine had no effects on in vitro angiogenesis using cultured HUVEC and NHDF. Ketamine can potentially be used as an anesthetic agent with no influence on angiogenesis. [ABSTRACT FROM AUTHOR]

Overactivation of the angiotensin-converting enzyme (ACE)/angiotensin II type 1 receptor (AT1R) pathway leads to vasoconstriction and elevated blood pressure. Persicaria minor (Huds.) Opiz is an herbal plant known for its antioxidant, anti-hyperlipidemic, and anti-atherosclerotic properties, with bioactive compounds that exhibit antihypertensive effects. Therefore, this study aimed to evaluate the antihypertensive effects of the standardized aqueous extract of P. minor leaf (AEPM) through the ACE/AT1R pathway in human umbilical vein endothelial cells (HUVECs) induced with phorbol 12-myristate 13-acetate (PMA). HUVECs were stimulated with PMA to induce ACE, with or without AEPM or captopril treatment, for 24 h. Subsequently, ACE mRNA expression, ACE protein levels, ACE activity, angiotensin II levels, and AT1R expression were measured. The results demonstrated that AEPM treatment significantly reduced ACE mRNA expression, ACE protein levels, ACE activity, angiotensin II levels, and AT1R expression in PMA-induced HUVECs. The modulatory effects of AEPM on the ACE/AT1R pathway were comparable to those of captopril. Ex vivo experiments further confirmed that AEPM reduced the contraction responses of rat aortic rings to PMA. In conclusion, P. minor effectively inhibits the ACE/AT1R pathway in PMA-induced HUVECs, suggesting its potential as a natural antihypertensive agent. [ABSTRACT FROM AUTHOR]

Introduction/objectives: JAK/STAT signaling inhibition exerts therapeutic effects on angiogenesis in rheumatoid arthritis (RA). However, whether the inhibitory effect differs among JAK inhibitors because of differing selectivity is unknown. Therefore, we compared the inhibitory effects of tofacitinib, baricitinib, peficitinib, upadacitinib, and filgotinib on angiogenesis. Method: RA-derived fibroblast-like synoviocytes (RA-FLS) were seeded on type I collagen gel, and human umbilical vein endothelial cells (HUVECs) were directly added. The control and aforementioned JAK inhibitors were added to the medium, followed by stimulation with interleukin (IL)-6 and soluble IL-6 receptor (sIL-6R). Each JAK inhibitor's concentration was determined based on estimated blood concentrations. The vascular endothelial growth factor (VEGF) concentration was evaluated with an enzyme-linked immunosorbent assay using the medium from the first exchange. A migration assay was performed, and HUVEC migration was evaluated using CD31 fluorescence immunostaining. Results: Hematoxylin–eosin staining showed that compared with the non-JAKi treatment group, the JAKi treatment group markedly degenerated in the sub-lining and deep lining, with decreased lymphocyte infiltration and neovascularization [Rooney's score subscale, non-JAKi vs JAKi (median, 6.5 vs 2.5, p = 0.005)]. In vitro, IL-6 and sIL-6R administration increased VEGF production from RA-FLS and promoted neovascularization in HUVECs, and JAK-inhibitor administration, which decreased VEGF production from RA-FLS and suppressed HUVEC migration, inhibited neovascularization in RA-FLS and HUVEC co-cultures. Conclusions: The JAK inhibitors suppressed IL-6-induced angiogenesis via decreased VEGF production and HUVEC migration in RA-FLS and HUVEC co-cultures. No significant differences were observed among the JAK inhibitors, whose anti-angiogenic effect may be an important mechanism for RA treatment. Key Points • JAK inhibitors inhibit angiogenesis in RA by reducing VEGF production from RA-derived fibroblast-like synoviocytes. • Our study provides new insights into RA treatment by elucidating the anti-angiogenic effect of JAK inhibitors. [ABSTRACT FROM AUTHOR]

Objective: To observe the protective effect and mechanism of hydroxyl safflower yellow A (HSYA) from myocardial ischemia-reperfusion injury on human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were treated with oxygen-glucose deprivation reperfusion (OGD/R) to simulate the ischemia reperfusion model, and cell counting kit-8 was used to detect the protective effect of different concentrations (1.25–160 µ mol/L) of HSYA on HUVECs after OGD/R. HSYA 80 µ mol/L was used for follow-up experiments. The contents of inflammatory cytokines interleukin (IL)-18, IL-1 β, monocyte chemotactic protein 1 (MCP-1), tumor necrosis factor α (TNF-α) and IL-6 before and after administration were measured by enzyme-linked immunosorbent assay. The protein expressions of toll-like receptor, NOD-like receptor containing pyrin domain 3 (NLRP3), gasdermin D (GSDMD) and GSDMD-N-terminal domain (GSDMD-N) before and after administration were detected by Western blot. NLRP3 inflammasome inhibitor cytokine release inhibitory drug 3 sodium salt (CRID3 sodium salt, also known as MCC950) and agonist were added, and the changes of NLRP3, cysteine-aspartic acid protease 1 (Caspase-1), GSDMD and GSDMD-N protein expressions were detected by Western blot. Results: HSYA inhibited OGD/R-induced inflammation and significantly decreased the contents of inflammatory cytokines IL-18, IL-1 β, MCP-1, TNF-α and IL-6 (P<0.01 or P<0.05). At the same time, by inhibiting NLRP3/Caspase-1/GSDMD pathway, HSYA can reduce the occurrence of pyroptosis after OGD/R and reduce the expression of NLRP3, Caspase-1, GSDMD and GSDMD-N proteins (P<0.01). Conclusions: The protective effect of HSYA on HUVECs after OGD/R is related to down-regulating the expression of NLRP3 inflammasome and inhibiting pyroptosis. [ABSTRACT FROM AUTHOR]

Objective To observe the effects of atorvastatin calcium on the cell proliferation and migration of human umbilical vein endothelial cells (HUVECs), and to screen the hub mRNA and co-related long non-coding RNA (lncRNA) of atorvastatin calcium on HUVECs. Methods HUVECs in the logarithmic growth phase were divided into the group 1 and group 2. Cells in the group 1 and group 2 were treated with 5 µmol/L atorvastatin calcium and the same amount of PBS for 24 h, respectively. ① Observation of effects of atorvastatin calcium on the cell proliferation and migration of HUVECs: The cell cycle was measured by flow cytometry, the cell proliferation was observed by CCK-8 method, and the cell migration was observed by scratch healing test. ② Screening of hub mRNAs and co-related lncRNAs in HUVECs treated with atorvastatin calcium: illumina platform was used to detect mRNA and lncRNA in cells of the two groups. Differentially expressed mRNAs and lncRNAs with |log2 Fold change| > 1. 5 and P adjusted < 0. 05 were screened using limma package in R Studio software. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were used to analyze the biological functions of differentially expressed mRNAs. The STRING database was used to analyze the protein interaction of differentially expressed mRNAs. The CytoNCA plug-in was used to screen the top 10 hub mRNAs with the highest mediation centrality, and the co-expressed lncRNAs with the hub mRNAs with Pearson correlation coefficient > 0. 99 were screened. Real-time fluorescence quantitative PCR (qPCR) was used to detect the hub mRNA and its coexpressed lncRNA in the two groups. Results Compared with the group 2, the proportion of cells in the G0/G1 phase of the group 1 was higher, the cell proliferation was inhibited, and the cell migration area was smaller after 24 h of culture (all P<0. 05) . Compared with the group 2, there were 1 510 differentially expressed mRNAs and 744 differentially expressed lncRNAs in the group 1; the biological functions of differentially expressed mRNAs involved 294 GO sets and 27 KEGG pathways; there were 10 hub mRNAs and 130 lncRNAs co-expressed with them, including AC017101. 1, AC125257. 1, AC004846. 1, etc. Compared with the group 2, the relative expression levels of HSP90AA1, CDK4, CCND1, PLK1, and PCNA in the group 1 were lower, and the relative expression levels of AC017101. 1, AC125257. 1, AC004846. 1, and AL390719. 1 were higher (all P<0. 05) . Conclusions Atorvastatin calcium can block the HUVECs in the G0/G1 phase and inhibit their proliferation and migration. There are 10 hub mRNAs such as HSP90AA1, CDK4 and CCND1, and 130 co-related lncRNAs such as AC017101. 1, AC125257. 1 and AC004846. 1 in atorvastatin calcium treatment of HUVECs. Atorvastatin calcium may promote the expression of co-related lncrnas such as AC017101. 1, AC125257. 1 and AC004846. 1 and down-regulate the expression of HSP90AA1, CDK4 and CCND1 mRNA in HUVECs. Thus, the cell cycle of HUVECs is arrested and cell proliferation and migration are inhibited. [ABSTRACT FROM AUTHOR]

Atherosclerosis is initiated by injury or damage to the vascular endothelial cell monolayer. Therefore, the early repair of the damaged vascular endothelium by a proliferation of neighbouring endothelial cells is important to prevent atherosclerosis and thrombotic events. Arthrospira platensis (AP) has been used as a dietary supplement, mainly due to its high content of vitamins, minerals, amino acids, and pigments such as chlorophylls, carotenoids, and phycocyanin, ingredients with antioxidant, anti-inflammatory, and anti-thrombotic properties. Therefore, in this prospective, placebo-controlled, data-driven, sample-size-estimated in vitro study, we tested whether an aqueous extract of AP at different concentrations (50, 100, and 200 µg/mL) had an effect on the different cellular parameters of human umbilical vein endothelial cells. Therefore, cell impedance measurement and cell proliferation were measured to investigate the monolayer formation. In addition, cell viability, integrity, and metabolism were analysed to evaluate singular cellular functions, especially the antithrombotic state. Furthermore, cell–cell and cell–substrate interactions were observed. The highest proliferation was achieved after the addition of 100 µg/mL. This was consistently confirmed by two independent optical experiments in cell cultures 48 h and 85 h after seeding and additionally by an indirect test. At this concentration, the activation or dysfunction of HUVECs was completely prevented, as confirmed by prostacyclin and interleukin-6 levels. In conclusion, in this study, AP induced a significant increase in HUVEC proliferation without inducing an inflammatory response but altered the hemostasiological balance in favour of prostacyclin over thromboxane, thereby creating an antithrombotic state. Thus, APE could be applied in the future as an accelerator of endothelial cell proliferation after, e.g., stent placement or atherosclerosis. [ABSTRACT FROM AUTHOR]

This study aims to demonstrate in vitro effects of osteopontin (OPN), an extracellular matrix molecule, on proliferation and migration in human umbilical vein endothelial cells (HUVEC) and Ishikawa human endometrial adenocarcinoma cells (Ishikawa) cultures. In addition, the effects of OPN on phosphatidylinositol 3 kinase (PI3K) and extracellular signal regulatory kinase 1/2 (ERK1/2), which are involved signaling pathways in cell cycle, were investigated. This research was carried out as a prospective cell culture study in Aydın Adnan Menderes University Biochemistry Laboratory. Two types of cell culture were used, HUVEC representing endothelial cells and Ishikawa representing tumour cells. The HUVEC and Ishikawa were extracted from liquid nitrogen and seeded in culture medium. With the increasing concentrations of recombinant human OPN (rhOPN) in both cell lines, proliferation and migration of cells and PI3K and ERK1/2 levels in the medium were investigated. Detection of cell proliferation in these two cell types with MTT test, and migration assay were performed. Proliferation was found to increase in both cell types with increasing levels of rhOPN (p < 0.01). In both cell cultures, rhOPN induced mesenchymal transformation, indicating migration at the highest (400 ng/mL) concentrations. Increasing levels of PI3K and ERK1/2 were measured with the increasing concentrations of rhOPN (p < 0.01 and p < 0.05, respectively). Proliferation, migration, and PI3K and ERK1/2 molecules are increased with rhOPN in both cell cultures studied. This suggests that OPN could be a candidate prognostic marker in endometrial cancer. [ABSTRACT FROM AUTHOR]

Introduction: Our research aimed to investigate the potential role and mechanism of lysyl oxidase (LOX)-like 2 (LOXL2) in atherosclerosis (AS) by using the human umbilical vein endothelial cells (HUVECs) stimulated by oxidized low-density lipoprotein (ox-LDL). Methods: HUVECs were treated with ox-LDL at different concentrations (0, 10, 25, 50, and 100 μg/mL) and incubated for 24 h. The transfection efficacy of siLOXL2 was investigated by Western blot and real-time quantitative polymerase chain reaction (RT-qPCR). Cell migration, intracellular ROS measurement, oxidative stress, enzyme-linked immunosorbent assay, and adhesion assays were carried out to examine the ox-LDL-induced HUVECs injury. RT-qPCR and Western blot were used to determine gene and protein expression levels. Results: LOXL2 protein expression increased in ox-LDL-induced endothelial cells (ECs). ox-LDL + siLOXL2 significantly inhibited the migration ability of HUVECs and reduced the expression of vascular endothelial growth factor A (VEGFA) and matrix metalloproteinase 9 gene expressions (all, p < 0.05). The ox-LDL + siLOXL2 significantly reduced intracellular ROS production and inhibited the expression of Malondialdehyde, whereas it markedly enhanced superoxide dismutase and catalase (all, p < 0.05). Supernatant levels of interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) were significantly attenuated by the ox-LDL + siLOXL2 treatment (all, p < 0.05). ox-LDL + siLOXL2 markedly suppressed the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 (all, p < 0.05). ox-LDL + siLOXL2 treatment remarkably reduced the expression of α-smooth muscle actin and vimentin, while increased CD31 and von Willebrand factor gene expression (all, p < 0.05). Conclusion: LOXL2 silencing is protected against ox-LDL-induced EC dysfunction, and the mechanism may be related to the inhibition of the EndMT pathway.

Abstract The aims of this study were to determine whether human umbilical cord mesenchymal stem cells (hucMSCs) modified by miRNA-25-3p (miR-25-3p) overexpression could promote venous endothelial cell proliferation and attenuate portal endothelial cell injury. HucMSCs and human umbilical vein endothelial cells (HUVEC) were isolated and cultured from human umbilical cord and characterized. Lentiviral vectors expressing miRNA-25-3p were transfected into hucMSCs and confirmed by PCR. We verified the effect of miR-25-3p-modified hucMSCs on HUVEC by cell co-culture and cell supernatant experiments. Subsequently, exosomes of miR-25-3p-modified hucMSCs were isolated from cell culture supernatants and characterized by WB, NTA and TEM. We verified the effects of miR-25-3p-modified exosomes derived from hucMSCs on HUVEC proliferation, migration, and angiogenesis by in vitro cellular function experiments. Meanwhile, we further examined the downstream target genes and signaling pathways potentially affected by miR-25-3p-modified hucMSC-derived exosomes in HUVEC. Finally, we established a rat portal vein venous thrombosis model by injecting CM-DiR-labeled hucMSCs intravenously into rats and examining the homing of cells in the portal vein by fluorescence microscopy. Histological and immunohistochemical experiments were used to examine the effects of miRNA-25-3p-modified hucMSCs on the proliferation and damage of portal vein endothelial cells. Primary hucMSCs and HUVECs were successfully isolated, cultured and characterized. Primary hucMSCs were modified with a lentiviral vector carrying miR-25-3p at MOI 80. Co-culture and cell supernatant intervention experiments showed that overexpression of miRNA-25-3p in hucMSCs enhanced HUVEC proliferation, migration and tube formation in vitro. We successfully isolated and characterized exosomes of miR-25-3p-modified hucMSCs, and exosome intervention experiments demonstrated that miR-25-3p-modified exosomes derived from hucMSCs similarly enhanced the proliferation, migration, and angiogenesis of HUVECs. Subsequent PCR and WB analyses indicated PTEN/KLF4/AKT/ERK1/2 as potential pathways of action. Analysis in a rat portal vein thrombosis model showed that miR-25-3p-modified hucMSCs could homing to damaged portal veins. Subsequent histological and immunohistochemical examinations demonstrated that intervention with miR-25-3p overexpression-modified hucMSCs significantly reduced damage and attenuated thrombosis in rat portal veins. The above findings indicate suggest that hucMSCs based on miR-25-3p modification may be a promising therapeutic approach for use in venous thrombotic diseases.

Introduction: Ischemic stroke (IS) is a global health concern, often tied to dyslipidemia and vascular endothelial dysfunction. MicroRNA-34a (miR-34a) was reported to be up-regulated in the blood samples of patients with IS, but the specific role of miR-34a and methylenetetrahydrofolate reductase (MTHFR) in IS remains to be elucidated. Methods: We studied 143 subjects: 71 IS patients, and 72 healthy controls. Human umbilical vein endothelial cells (HUVECs) were cultured and transfected with a miR-34a mimic, inhibitor, or negative control. The miR-34a expression in serum and HUVECs was quantified via quantitative reverse transcription polymerase chain reaction (qRT-PCR). Viability and apoptosis of HUVECs were assessed using CCK-8 assay and flow cytometry. The expression levels of bcl-2, bax, cyt-c, cleaved caspase 3, MTHFR, and homocysteine were measured by Western blot or enzyme-linked immunosorbent assay (ELISA). The relationship between miR-34a and MTHFR was verified by luciferase reporter assay. The levels of MTHFR and homocysteine in serum were examined by ELISA. Results: MiR-34a expression was increased in IS patients and inhibited viability of HUVECs while promoting their apoptosis. Overexpression of miR-34a up-regulated pro-apoptotic proteins (bax, cyt-c and cleaved caspase 3) and down-regulated anti-apoptotic protein bcl-2 in HUVECs. MTHFR was identified as the downstream target of miR-34a and its expression was reduced by miR-34a overexpression, while homocysteine levels increased. Consistently, MTHFR levels were lower and homocysteine levels were higher in IS patients compared with controls. Discussion: Our results suggest that up-regulated miR-34a plays a role in the pathogenesis of IS, potentially through inhibiting MTHFR expression and increasing homocysteine in endothelial cells. Therefore, miR-34a might be a therapeutic target for IS [ABSTRACT FROM AUTHOR]

Diabetes mellitus cases and complications, including difficult-to-heal wounds, are increasing worldwide. Diabetic wounds frequently exhibit impaired angiogenesis due to hyperglycemia, which disrupts many factors that promote angiogenesis, including the release of vascular endothelial growth factor (VEGF). Physalis angulata L, an herb in the Solanaceae family, is used as a herbal medicine for many ailments, such as treating wounds. The fruit of P. angulata is rich in lupeol, a compound that can enhance wound healing by upregulating the expression of certain growth factors. This study aimed to determine the effect of P. angulata fruit extract on endothelial cell migration and VEGF concentration in wound healing in hyperglycemic endothelial cell cultures. Human umbilical vein endothelial cells (HUVECs) were exposed to high glucose (22 mM/dL) for 6 and 24 h with or without 0.025%, 0.05%, and 0.1% concentrations of P. angulata fruit extract. Cell migration was examined in HUVECs by scratching and measuring the distance. VEGF concentration was measured using enzyme-linked immunosorbent assay methods. P. angulata fruit extract at a concentration of 0.025% for 24 h significantly stimulated cell migration and VEGF concentration in HUVECs exposed to hyperglycemia. P. angulata fruit extract is a promising herb for treating diabetic wounds. [ABSTRACT FROM AUTHOR]

The aims of this study were to determine whether human umbilical cord mesenchymal stem cells (hucMSCs) modified by miRNA-25-3p (miR-25-3p) overexpression could promote venous endothelial cell proliferation and attenuate portal endothelial cell injury. HucMSCs and human umbilical vein endothelial cells (HUVEC) were isolated and cultured from human umbilical cord and characterized. Lentiviral vectors expressing miRNA-25-3p were transfected into hucMSCs and confirmed by PCR. We verified the effect of miR-25-3p-modified hucMSCs on HUVEC by cell co-culture and cell supernatant experiments. Subsequently, exosomes of miR-25-3p-modified hucMSCs were isolated from cell culture supernatants and characterized by WB, NTA and TEM. We verified the effects of miR-25-3p-modified exosomes derived from hucMSCs on HUVEC proliferation, migration, and angiogenesis by in vitro cellular function experiments. Meanwhile, we further examined the downstream target genes and signaling pathways potentially affected by miR-25-3p-modified hucMSC-derived exosomes in HUVEC. Finally, we established a rat portal vein venous thrombosis model by injecting CM-DiR-labeled hucMSCs intravenously into rats and examining the homing of cells in the portal vein by fluorescence microscopy. Histological and immunohistochemical experiments were used to examine the effects of miRNA-25-3p-modified hucMSCs on the proliferation and damage of portal vein endothelial cells. Primary hucMSCs and HUVECs were successfully isolated, cultured and characterized. Primary hucMSCs were modified with a lentiviral vector carrying miR-25-3p at MOI 80. Co-culture and cell supernatant intervention experiments showed that overexpression of miRNA-25-3p in hucMSCs enhanced HUVEC proliferation, migration and tube formation in vitro. We successfully isolated and characterized exosomes of miR-25-3p-modified hucMSCs, and exosome intervention experiments demonstrated that miR-25-3p-modified exosomes derived from hucMSCs similarly enhanced the proliferation, migration, and angiogenesis of HUVECs. Subsequent PCR and WB analyses indicated PTEN/KLF4/AKT/ERK1/2 as potential pathways of action. Analysis in a rat portal vein thrombosis model showed that miR-25-3p-modified hucMSCs could homing to damaged portal veins. Subsequent histological and immunohistochemical examinations demonstrated that intervention with miR-25-3p overexpression-modified hucMSCs significantly reduced damage and attenuated thrombosis in rat portal veins. The above findings indicate suggest that hucMSCs based on miR-25-3p modification may be a promising therapeutic approach for use in venous thrombotic diseases. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression and significance of the mechanosensitive ion channel Piezol in human placental chorionic arteries (HPCA) of pre-eclampsia (PE) patients. Methods HPCA tissues were collected from pregnant women undergoing cesarean section delivery at the Department of Obstetrics, Affiliated Hospital of South-west Medical University. The samples included a normal control (NC) group, a PE group, and a severe pre eclampsia (SPE) group, each comprising 10 cases. Immunofluorescence staining was used to detect the expression of Piezol and the endothelial marker CD31 in the HPCA of the NC, PE, and SPE groups. Real-time quantitative PCR (qPCR) was employed to measure the relative mRNA expression of Piezol in the HPCA of the NC, PE, and SPE groups. Western blotting (WB) was conducted to assess the relative protein expression of Piezol in the HPCA of the NC, PE, and SPE groups. Hydrogen peroxide (H2O2) was used to simulate oxidative stress in human umbilical vein endothelial cells (HUVECs), and qPCR and WB were performed to evaluate the mRNA and protein expression of Piezol in the NC group, H2O2, - trea- ted 6 h group, and H2O2 treated 12 h group of HUVECs. Results Piezol co localized with the endothelial marker CD31 in the HPCA tissues of the NC, PE, and SPE groups. The mRNA expression of Piezol showed a decreasing trend in the HPCA tissues of the NC, PE, and SPE groups, with statistically significant differences (P<0.05). Similarly, the protein expression of Piezol exhibited a decreasing trend in the HPCA tissues of the NC, PE, and SPE groups, with statistically significant differences (P<0.05). In HUVECs, both mRNA and protein expression of Piezol showed a decreasing trend in the NC group, H2O2 treated 6 h group, and H2O2 treated 12h group, with statistically significant differences (P<0.05). Conclusion Piezol is expressed in endothelial cells of HPCA tissues, and its abnormal expression may be associated with the pathophysiological changes in pre-eclampsia. The occurrence and development of pre-eclampsia related oxidative stress response may lead to decreased expression of Piezol. [ABSTRACT FROM AUTHOR]

HUVECs in the experimental group were treated with 50, 100 and 200 mg/L chrysotile for 24, 48 and 72 h. The control group was cultured with RPMI 1640 medium only. Morphological changes of HUVECs were observed. Cell viability was detected by MTT assay. β-Galactosidase staining was used to analyze the cell senescence. The cell survival rate was determined by tetramethylazolium blue assay. Wnt5a, p16 and p21 expression were investigated by qRT-PCR. Results Cells of the experimental group were oval and irregular, and some of them were round or irregular, with naked nuclei and vacuolar, and dead cells were present. With the increase of chrysotile concentration and exposure time, the level of cell viability decreased and the number of aged cells increased gradually (P＜0.05). The cell growth of HUVECs was more active when 100 mg/L chrysotile treated for 24 h. Compared with the control group, the mRNA expression levels of Wnt5a, p16 and p21 were increased in the experimental group (P＜0.05). Conclusion Chrysotile can promote HUVECs senescence, and Wnt5a, p16 and p21 are involved in this mechanism. [ABSTRACT FROM AUTHOR]

Exposure to microgravity during spaceflight induces the alterations in endothelial cell function associated with post-flight cardiovascular deconditioning. PIEZO1 is a major mechanosensitive ion channel that regulates endothelial cell function. In this study, we used a two-dimensional clinostat to investigate the expression of PIEZO1 and its regulatory mechanism on human umbilical vein endothelial cells (HUVECs) under simulated microgravity. Utilizing quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis, we observed that PIEZO1 expression was significantly increased in response to simulated microgravity. Moreover, we found microgravity promoted endothelial cells migration by increasing expression of PIEZO1. Proteomics analysis highlighted the importance of C-X-C chemokine receptor type 4(CXCR4) as a main target molecule of PIEZO1 in HUVECs. CXCR4 protein level was increased with simulated microgravity and decreased with PIEZO1 knock down. The mechanistic study showed that PIEZO1 enhances CXCR4 expression via Ca2+ influx. In addition, CXCR4 could promote endothelial cell migration under simulated microgravity. Taken together, these results suggest that the upregulation of PIEZO1 in response to simulated microgravity regulates endothelial cell migration due to enhancing CXCR4 expression via Ca2+ influx. [ABSTRACT FROM AUTHOR]

Objectives: Deep-venous thrombosis (DVT) refers to abnormal blood clotting in the deep vein cavity, and post-thrombotic syndrome (PTS) is the most frequent complication. The study explored the impact of microRNA 181b-5p on DVT progression based on human umbilical vein endothelial cells (HUVECs).Methods: Levels of miR-181b-5p were examined in 150 cases with acute lower extremity DVT. ROC curve and K-M plot were drawn for clinical value assessment. The role of miR-181b-5p in HUVECs viability, migration, apoptosis, inflammatory response and adhesion factors’ release was investigated. Target gene of miR-181b-5p was predicted, and its role in cell function was explored.Results: Low-expressed miR-181b-5p showed favorable diagnostic performance in differentiating DVT with the AUC of 0.948. Patients with low miR-181b-5p had a high incidence of PTS. miR-181b-5p overexpression promoted HUVECs’ viability and migration, while inhibiting cell apoptosis and release of inflammatory and adhesion cytokines. As the target gene of miR-181b-5p, HEY2 overexpression reversed the role of miR-181b-5p in HUVECs.Conclusion: MiR-181b-5p serves as a potential biomarker for DVT diagnosis and PTS development. Overexpression of this miRNA targeted HEY2 to alleviate endothelial cell damage.

Atherosclerosis is characterized by accumulation of lipid that triggers arterial inflammation. This study aims to investigate the protective effect of alfalfa flavonoid extract (AFE) on HUVEC cells treated with lipopolysaccharide (LPS) and rats fed with high-fat diet (HFD). AFE reduced the expression of proinflammatory factors (COX-2, iNOS, IL-1β, IL-6 and TNF-α), intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, and inhibited activation of NF-κB and MAPK pathways. AFE alleviated vascular endothelial lesions by reducing HFD-enhanced levels of triglyceride, total cholesterol and low-density lipoprotein but increased high-density lipoprotein. Mechanically, AFE ameliorated HFD-enhanced liver steatosis through downregulating gene expression for lipid biosynthesis (SREBP2, HMGCR and FASN) and lipid uptake (LDLR), and through upregulating CYP7A1 expression for lipid catabolism and excretion. These data suggest that AFE possesses the anti-atherosclerotic potential through controlling inflammation, hyperlipidemia and lipid metabolism, and can be used as a functional food additive to benefit patients with atherosclerosis.

Vascularization is a key step to achieve pulp tissue regeneration and in vitro pre-vascularized dental pulp tissue could be applied as a graft substitute for dental pulp tissue repair. In this study, human dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (hUVECs) were co-cultured in 3D Matrigel and 150 mV/mm electric fields (EFs) were used to promote the construction of pre-vascularized dental pulp tissue. After optimizing co-cultured ratio of two cell types, immunofluorescence staining, and live/dead detection were used to investigate the effect of EFs on cell survival, differentiation and vessel formation in 3D engineered dental pulp tissue. RNA sequencing was used to investigate the potential molecular mechanisms by which EF regulates vessel formation in 3D engineered dental pulp tissue. Here we identified that EF-induced pre-vascularized engineered dental pulp tissue not only had odontoblasts, but also had a rich vascular network, and smooth muscle-like cells appeared around the blood vessels. The GO enrichment analysis showed that these genes were significantly enriched in regulation of angiogenesis, cell migration and motility. The most significant term of the KEGG pathway analysis were NOTCH signaling pathway and Calcium signaling pathway etc. The PPI network revealed that NOTCH1 and IL-6 were central hub genes. Our study indicated that EFs significantly promoted the maturation and stable of blood vessel in 3D engineered pulp tissue and provided an experimental basis for the application of EF in dental pulp angiogenesis and regeneration.

Abstract Bioabsorbable sutures can improve the medical functions of existing non-absorbable sutures, and may produce new medical effects, and are expected to become a new generation of medical degradable materials. In this study, the cytocompatibility of triclosan coated polyglactin910 sutures (CTS-PLGA910) was analyzed and different concentrations of sutures were prepared. The effects of sutures on the cytotoxicity and cell proliferation of HUVEC were studied by CCK-8 assay. The hemolysis, total antioxidant capacity (T-AOC) activity and nitric oxide (NO) content were investigated to improve the blood compatibility of sutures. The results showed that the hemolysis rate of CTS-PLGA910 was less than 5%. After treatment on HUVEC cells for 48 and 72 h, there was no significant change in NO content in CTS-PLGA910 groups compared with the control group, while T-AOC activity and antioxidant capacity were significantly increased in medium and high dose groups. In summary, the blood compatibility and cell compatibility were significantly improved, which provided a basis for the clinical application of sutures in the future. Graphical Abstract

To explored the protective effect and mechanism of Chinese yam total protein in human umbilical vein endothelial cells (HUVECs) with high-concentration D-glucose (HG)-induced oxidative stress injury. The HUVECs injury model was established by 50 mmol/L HG. HUVECs were randomly divided into control, model, low-dose Chinese yam total protein (0.5 mg/mL), and high-dose Chinese yam total protein (1 mg/mL) groups. The effects of Chinese yam total protein on the viability, morphology, and angiogenetic ability of HUVECs treated with HG were evaluated. The reactive oxygen species (ROS) level, superoxide dismutase (SOD) and catalase (CAT) activities in cells were determined. The nitric oxide (NO), 8-hydroxy deoxyguanosine (8-OHdG), malondialdehyde (MDA), interleukin-1β (IL-1β), and interleukin-6 (IL-6) levels in culture medium supernatant were measured. Protein expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X (Bax), thioredoxin-interacting protein (Txnip), NOD-like receptor protein 3 (NLRP3), cleaved Caspase-1, and IL-1β were determined. The results showed that Chinese yam total protein could significantly increased the cell viability of HUVECs treated with HG (P

In this work, the magnetron sputtering method is used to deposit tantalum-containing coatings on NiTi shape memory alloys under various sputtering atmospheres including (i) 100% argon, (ii) 16.6% oxygen and 83.4% argon, and (iii) 16.6% nitrogen and 83.4% argon. This research examines how the sputtering atmosphere can impact surface morphology, roughness, wettability, biocompatibility, in vitro cytocompatibility, and antibacterial performance of tantalum-containing coatings. The calculation of nickel ions release by coupled inductively coupled plasma-optical emission spectrometry displayed that none of the coated samples showed any traces of nickel, unlike the bare NiTi substrate. This has led to an improvement in biocompatibility. Topographical measurements demonstrated the formation of a smooth surface under the argon-oxygen atmosphere, whereas the deposited layer in the argon-nitrogen atmosphere exhibited irregular ups and downs. The coatings deposited under the argon-nitrogen atmosphere result in a hydrophobic surface with many hills and valleys which can considerably enhance protein and cellular absorption. Besides, the samples coated under an argon-oxygen atmosphere with a smooth surface exhibited excellent antibacterial efficiency only as early as 6 h of incubation. This is probably due to the formation of amorphous tantalum pentoxide.

Somatic activating mutations of PIK3CA are associated with development of vascular malformations (VMs). Here, we describe a microfluidic model of PIK3CA-driven VMs consisting of human umbilical vein endothelial cells expressing PIK3CA activating mutations embedded in three-dimensional hydrogels. We observed enlarged, irregular vessel phenotypes and the formation of cyst-like structures consistent with clinical signatures and not previously observed in cell culture models. Pathologic morphologies occurred concomitant with up-regulation of Rac1/p21-activated kinase (PAK), mitogen-activated protein kinase cascades (MEK/ERK), and mammalian target of rapamycin (mTORC1/2) signaling networks. We observed differential effects between alpelisib, a PIK3CA inhibitor, and rapamycin, an mTORC1 inhibitor, in mitigating matrix degradation and network topology. While both were effective in preventing vessel enlargement, rapamycin failed to reduce MEK/ERK and mTORC2 activity and resulted in hyperbranching, while inhibiting PAK, MEK1/2, and mTORC1/2 mitigates abnormal growth and vascular dilation. Collectively, these findings demonstrate an in vitro platform for VMs and establish a role of dysregulated Rac1/PAK and mTORC1/2 signaling in PIK3CA-driven VMs.

Objective To explore the changes in morphology, proliferation activity and VE-cadherin, E-cadherin, and N-cadherin expression of human umbilical vein endothelial cells (HUVECs） under different concentrations of glucose. Methods HUVECs were divided into the normal control group (glucose concentration of 5. 5 mmol/L）, mannitol group (glucose concentration of 5. 5 mmol/L + mannitol concentration of 24. 5 mmol/L）, and high glucose group (glucose concentration of 11. 1，22. 2, and 33. 3 mmol/L, respectively）, and were cultured for 24，48, and 72 h. Cell morphology was observed under the microscope, and cell proliferation in each group was detected by CCK-8 reagent. Western blotting was used to detect the expression levels of VE-cadherin, E-cadherin, and N-cadherin in HUVECs in each group. Results After 72 h of cultivation, the cells in the normal control group showed a quasi-circular shape, while cells in the high glucose group and mannitol group gradually changed from circular and elliptical to cord shape, with a decrease in cell number and an increase in cell gaps compared with those before. After 24 h of cultivation, the cell proliferation rate of the high glucose group was higher than that of the normal control group. With the glucose concentration increased, the cell proliferation rate of the high glucose group showed a trend of first increasing and then decreasing. After 48 and 72 h of cultivation, the cell proliferation rate of the high glucose group was lower than that of the normal control group. With the glucose concentration increased, the cell proliferation rate of the high glucose group gradually decreased. Statistically significant differences were found in the cell proliferation rates among the above groups (all P<0. 05）. After 24 and 72 h of cultivation, compared with the normal control group, the expression levels of N-cadherin, VE-cadherin, and E-cadherin in HUVECs increased in the high glucose group (all P<0. 05）； and with the increase of glucose concentrations, the N-cadherin, VE-cadherin, and E-cadherin of cells in the high glucose group showed a trend of first increasing and then decreasing. Conclusion Under high glucose conditions, HUVECs undergo morphological transformation, with a decrease in proliferation rate, accompanied by changes in the expression levels of N-cadherin, VE-cadherin, and E-cadherin. [ABSTRACT FROM AUTHOR]

To explored the protective effect and mechanism of Chinese yam total protein in human umbilical vein endothelial cells (HUVECs) with high-concentration D-glucose (HG)-induced oxidative stress injury. The HUVECs injury model was established by 50 mmol/L HG. HUVECs were randomly divided into control, model, low-dose Chinese yam total protein (0.5 mg/mL), and high-dose Chinese yam total protein (1 mg/mL) groups. The effects of Chinese yam total protein on the viability, morphology, and angiogenetic ability of HUVECs treated with HG were evaluated. The reactive oxygen species (ROS) level, superoxide dismutase (SOD) and catalase (CAT) activities in cells were determined. The nitric oxide (NO), 8-hydroxy deoxyguanosine (8-OHdG), malondialdehyde (MDA), interleukin-1β (IL-1β), and interleukin-6 (IL-6) levels in culture medium supernatant were measured. Protein expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X (Bax), thioredoxin-interacting protein (Txnip), NOD-like receptor protein 3 (NLRP3), cleaved Caspase-1, and IL-1β were determined. The results showed that Chinese yam total protein could significantly increased the cell viability of HUVECs treated with HG (P<0.01), improved cell apoptotic morphology, and significantly increased the activities of SOD and CAT (P<0.05, P<0.01) and ratio of apoptosis-related proteins (Bcl-2/Bax), angiogenetic ability and nitric oxide (NO) secretion. Moreover, Chinese yam total protein significantly decreased the levels of ROS, MDA, 8-OHdG, IL-1β, and IL-6 (P<0.05, P<0.01) and the expression levels of inflammation-related proteins (Txnip, NLRP3, cleaved Caspase-1, and IL-1β) (P<0.05, P<0.01). Changes in these indicators showed a significant concentration-dependent manner. The results indicate that Chinese yam total protein may protect against HG-induced oxidative stress by inhibiting the Txnip/NLRP3 signaling pathway, restoring the oxidative balance, and reducing the inflammatory response of HUVECs. [ABSTRACT FROM AUTHOR]

Stem cell spheroid is a promising graft substitute for bone tissue engineering. Spheroids obtained by 3D culture of STRO1+ Gingival Mesenchymal Stem Cells (sGMSCs) (sGMSC spheroids, GS) seldom express angiogenic factors, limiting their angiogenic differentiation in vivo. This study introduced a novel stem cell spheroid with osteogenic and angiogenic potential through 3D co-culture of sGMSCs and Human Umbilical Vein Endothelial Cells (HUVECs) (sGMSC/HUVEC spheroids, GHS). GHS with varying seeding ratios of sGMSCs to HUVECs (GHR) were developed. Cell fusion within the GHS system was observed via immunofluorescence. Calcein-AM/PI staining and chemiluminescence assay indicated cellular viability within the GHS. Furthermore, osteogenic and angiogenic markers, including ALP, OCN, RUNX2, CD31, and VEGFA, were quantified and compared with the control group comprising solely of sGMSCs (GS). Incorporating HUVECs into GHS extended cell viability and stability, initiated the expression of angiogenic factors CD31 and VEGFA, and upregulated the expression of osteogenic factors ALP, OCN, and RUNX2, especially when GHS with a GHR of 1:1. Taken together, GHS, derived from the 3D co-culture of sGMSCs and HUVECs, enhanced osteogenic and angiogenic capacities in vitro, extending the application of cell therapy in bone tissue engineering. [ABSTRACT FROM AUTHOR]

This document is a list of references for a research article on the topic of endoplasmic reticulum stress and endothelial dysfunction. The references include various studies and articles that explore the relationship between these two factors and their implications for conditions such as diabetes and cardiovascular disease. [Extracted from the article]

Hyperglycaemia is a key factor in the progression of diabetes complications. Dapagliflozin (DAPA), a new type of hypoglycaemic agent, has been shown to play an important role in anti‐apoptotic, anti‐inflammatory and antioxidant activities. Previous studies have demonstrated an endothelial protective effect of DAPA, but the underlying mechanism was still unclear. Autophagy is a homeostatic cellular mechanism that circulates unfolded proteins and damaged organelles through lysosomal dependent degradation. In this study, we aimed to investigate whether DAPA plays a protective role against high glucose (HG)‐induced endothelial injury through regulating autophagy. The results showed that DAPA treatment resulted in increased cell viability. Additionally, DAPA treatment decreased interleukin (IL)‐1β, IL‐6, and tumour necrosis factor‐α levels in endothelial cells subjected to HG conditions. We observed that HG inhibited autophagy, and DAPA increased the autophagy level by inhibiting the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signalling pathway. Chloroquine reversed all of these beneficial effects as an autophagy inhibitor. In summary, the endothelial protective effect of DAPA in HG can be attributed in part to its role in activating of autophagy via the AKT/mTOR signalling pathway. Therefore, suggesting that the activation of autophagy by DAPA may be a novel target for the treatment of HG‐induced endothelial cell injury. [ABSTRACT FROM AUTHOR]

Jinzhao Liu,1,2,&ast; Jia Dong,1,&ast; Xibo Pei1,2 1State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, 610041, People’s Republic of China; 2Department of Prosthodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, 610041, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jia Dong; Xibo Pei, Email scu2013dongjia@sina.com; xbpei@hotmail.comPurpose: The promotion of angiogenesis is an effective strategy for skin wound repair. While the transplantation of endothelial cells has shown promise in vascularization, the underlying mechanism remains unclear. Recent studies have suggested that transplanted cells undergo apoptosis in a short period and release apoptotic extracellular vesicles (ApoEVs) that may have therapeutic potential.Methods: In this study, we isolated ApoEVs from human umbilical vein endothelial cells (HUVECs) and characterized their properties. In vitro, we assessed the effects of ApoEVs on the proliferation, migration, and differentiation of endothelial cells and fibroblasts. In vivo, we investigated the therapeutic role of ApoEVs-AT in full-thickness skin wounds, evaluating wound closure rate, re-epithelialization, granulation tissue formation, vascularization, scar area, and collagen 3(Col3)/collagen 1(Col 1) ratio.Results: ApoEVs derived from HUVECs displayed typical characteristics. In vitro, ApoEVs significantly enhanced the proliferation, migration, tube formation, and expression of angiogenic-related genes in endothelial cells and slightly promoted the proliferation and migration of fibroblasts. In vivo, ApoEVs improved the wound closure rate, re-epithelialization, the formation of granulation tissue, and vascularization. Besides, ApoEVs reduced scar formation, accompanied by an increase in the Col 3/ Col 1 ratio.Conclusion: Given their abundant source and effectiveness, this study provided a novel approach for angiogenesis in tissue regeneration and deepened the understanding of from death to regeneration.Keywords: apoptotic extracellular vesicles, human umbilical vein endothelial cells, skin regeneration, angiogenesis

Abstract Background Diabetes mellitus (DM) and its associated vascular complications have become a worldwide health concern. The effects and mechanism of vitamin D supplementation on endothelial function under high glucose condition remain elusive. Methods Human umbilical vein endothelial cells (HUVECs) were treated with 35 mM glucose, then 100 nM vitamin D were added. Transwell migration assay, CCK-8, immunofluorescence, flow cytometry, autophagy flux and transmission electric microscope were performed. Results Vitamin D reduced apoptosis, promoted migration and enhanced viability of HUVECs, decreased TIPE1 (Tumor necrosis factor-α-induced protein 8-like 1) under high glucose conditions. Overexpression of TIPE1 reverses the effects of vitamin D by increasing ROS production, inflammation, cell apoptosis, and suppressing autophagy, cell migration and viability. And vitamin D negatively correlated with TIPE1 mRNA level in DM patients. Conclusions Vitamin D reverses the harmful effects of high glucose on HUVECs by reducing TIPE1 expression. And vitamin D supplementation could help to alleviate high glucose-induced injury in type 2 diabetes mellitus patients with microvascular complications.

Overactivation of the angiotensin-converting enzyme (ACE)/angiotensin II type 1 receptor (AT1R) pathway leads to vasoconstriction and elevated blood pressure. Persicaria minor (Huds.) Opiz is an herbal plant known for its antioxidant, anti-hyperlipidemic, and anti-atherosclerotic properties, with bioactive compounds that exhibit antihypertensive effects. Therefore, this study aimed to evaluate the antihypertensive effects of the standardized aqueous extract of P. minor leaf (AEPM) through the ACE/AT1R pathway in human umbilical vein endothelial cells (HUVECs) induced with phorbol 12-myristate 13-acetate (PMA). HUVECs were stimulated with PMA to induce ACE, with or without AEPM or captopril treatment, for 24 h. Subsequently, ACE mRNA expression, ACE protein levels, ACE activity, angiotensin II levels, and AT1R expression were measured. The results demonstrated that AEPM treatment significantly reduced ACE mRNA expression, ACE protein levels, ACE activity, angiotensin II levels, and AT1R expression in PMA-induced HUVECs. The modulatory effects of AEPM on the ACE/AT1R pathway were comparable to those of captopril. Ex vivo experiments further confirmed that AEPM reduced the contraction responses of rat aortic rings to PMA. In conclusion, P. minor effectively inhibits the ACE/AT1R pathway in PMA-induced HUVECs, suggesting its potential as a natural antihypertensive agent.

Atherosclerosis is initiated by injury or damage to the vascular endothelial cell monolayer. Therefore, the early repair of the damaged vascular endothelium by a proliferation of neighbouring endothelial cells is important to prevent atherosclerosis and thrombotic events. Arthrospira platensis (AP) has been used as a dietary supplement, mainly due to its high content of vitamins, minerals, amino acids, and pigments such as chlorophylls, carotenoids, and phycocyanin, ingredients with antioxidant, anti-inflammatory, and anti-thrombotic properties. Therefore, in this prospective, placebo-controlled, data-driven, sample-size-estimated in vitro study, we tested whether an aqueous extract of AP at different concentrations (50, 100, and 200 µg/mL) had an effect on the different cellular parameters of human umbilical vein endothelial cells. Therefore, cell impedance measurement and cell proliferation were measured to investigate the monolayer formation. In addition, cell viability, integrity, and metabolism were analysed to evaluate singular cellular functions, especially the antithrombotic state. Furthermore, cell–cell and cell–substrate interactions were observed. The highest proliferation was achieved after the addition of 100 µg/mL. This was consistently confirmed by two independent optical experiments in cell cultures 48 h and 85 h after seeding and additionally by an indirect test. At this concentration, the activation or dysfunction of HUVECs was completely prevented, as confirmed by prostacyclin and interleukin-6 levels. In conclusion, in this study, AP induced a significant increase in HUVEC proliferation without inducing an inflammatory response but altered the hemostasiological balance in favour of prostacyclin over thromboxane, thereby creating an antithrombotic state. Thus, APE could be applied in the future as an accelerator of endothelial cell proliferation after, e.g., stent placement or atherosclerosis.

Background Damage to the coronary arteries during the acute phase of Kawasaki disease (KD) is linked to inflammatory cell infiltration, myointimal proliferation, and endothelial cell (EC) dysfunction. To understand the response of ECs to KD treatment, we studied the genome-wide transcriptional changes in cultured ECs incubated with KD sera before and after treatment with or without atorvastatin. Methods and Results RNA sequencing of human umbilical vein ECs incubated with pooled sera from patients with acute KD before or after treatment with intravenous immunoglobulin and infliximab revealed differentially expressed genes in interleukin-1, tumor necrosis factor-α, and inflammatory cell recruitment pathways. Subacute sera pooled from patients treated with intravenous immunoglobulin, infliximab, and atorvastatin uniquely induced expression of NOS3, Kruppel like factor (KLF2, and KLF4 (promotes EC homeostasis and angiogenesis) and ZFP36 ring finger protein (ZFP36) and suppressor of cytokine signaling 3 (SOCS3) (suppresses inflammation), and suppressed expression of TGFB2 and DKK1 (induces endothelial-mesenchymal transition) and sphingosine kinase 1 (SPHK1) and C-X-C motif chemokine ligand 8 (CXCL8) (induces inflammation). Conclusions These results suggest that atorvastatin treatment of patients with acute KD may improve EC health, reduce mediators of inflammation produced by ECs, and block KD-induced myofibroblast proliferation.

Angiotensin-converting enzyme (ACE) plays a crucial role in the pathogenesis of hypertension. Piper sarmentosum Roxb., an herb known for its antihypertensive effect, lacks a comprehensive understanding of the mechanism underlying its antihypertensive action. This study aimed to elucidate the antihypertensive mechanism of aqueous extract of P. sarmentosum leaves (AEPS) via its modulation of the ACE pathway in phorbol 12-myristate-13-acetate (PMA)-induced human umbilical vein endothelial cells (HUVECs). HUVECs were divided into five groups: control, treatment with 200 µg/mL AEPS, induction 200 nM PMA, concomitant treatment with 200 nM PMA and 200 µg/mL AEPS, and treatment with 200 nM PMA and 0.06 μM captopril. Subsequently, ACE mRNA expression, protein level and activity, angiotensin II (Ang II) levels, and angiotensin II type 1 receptor (AT1R) and angiotensin II type 2 receptor (AT2R) mRNA expression in HUVECs were determined. AEPS successfully inhibited ACE mRNA expression, protein and activity, and angiotensin II levels in PMA-induced HUVECs. Additionally, AT1R expression was downregulated, whereas AT2R expression was upregulated. In conclusion, AEPS reduces the levels of ACE mRNA, protein and activity, Ang II, and AT1R expression in PMA-induced HUVECs. Thus, AEPS has the potential to be developed as an ACE inhibitor in the future. [ABSTRACT FROM AUTHOR]

Objective We aimed to (i) investigate the protective effect of active components of water extract of huangqi (milkvetch root) on high glucose (25 mmol/L) induced injury of human umbilical vein endothelial cells (HUVEC) and (ii) elucidate the underlying mechanism. Methods Male SD rats were continuously fed with water extract [20 g/(kg·d)] for 7 d, and blood samples were colleeted 1, 2 and 4 h after the last gavage, combined with supernatant, and performed ultra-performance liquid chromatography (UPLC-MS) analysis to screen the active components of the extract in rats. Cells were divided into normal group, model group, different active components groups treated with different concentrations, and the concerted application of active components group. Cell viability was measured by Cell Counting Kit-8 to explore the optimal concentration of the effective components of milkvetch root. Oxidative damage was measured by reactive oxygen species (ROS). The enzyme-linked immunosorbent assay measured the contents of superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA); the protein expression level of nuclear factor E2 related factor (Nrf 2) and heme oxygenase 1 (HO-1) by Western blotting. Results Formononetin, daidzein, and calycosin are the active components of milkvetch root water extract.Compared with the normal group, the cell viability of the model group was increased (P<0.05). Compared with the model group, cell viability was decreased in each concentration(26, 13, 6.5 μmol/L) of formononetin group (P<0.05), a high dose of daidzein (0.11 μmol/L) could reduce cell viability (P<0.05). Compared with the normal group, the ROS levels in the model group were increased(P<0.05); compared with the model group, the ROS levels in the formononetin high and medium concentrations(26, 13 μmol/L) groups, different concentrations of daidzein (0.11, 0.06, 0.03 μmol/L), the calycosin high concentration group (7.00 μmol/L) were decreased (P<0.05). The combined active components inhibitive rate was higher than the single active components, and highest in the high concentration group. Compared with the normal group, Nrf2 and HO-1 protein expressions were decreased in the model group (P<0.05); compared with the model group, Nrf2 and HO-1 proteins were increased in the group with high concentration of concerted application group (P<0.05). Compared with the normal group, SOD, CAT were decreased (P<0.05), MDA increased (P<0.05) in the model group; compared with the model group, SOD, CAT in the high concentration concerted application group were increased (P<0.05), MDA was significantly decreased (P<0.05). Conclusion The active components of milkvetch root water extract, formononetin, daidzein, calycosin can protect HUVEC from high glucose-induced injury. The mechanism may be related to the regulation of Nrf2, HO-1, SOD, CAT, and MDA expression and their antioxidant effects. [ABSTRACT FROM AUTHOR]

Background: Diabetic foot ulcer (DFU) carries high rates of major amputation and mortality. Aims: The goals of this study were to identify expression of circulating lncRNA DLEU1 and miR-96-5p in patients with diabetic foot ulcer (DFU) and to explore the function of lncRNA DLEU1/miR-96-5p axis in DFU. Methods: Matched patients with DFU and healthy individuals were randomly selected. Serum samples from all subjects were used for circulating lncRNA DLEU1 and miR-96-5p assessment by RT-qPCR. Receiver operating characteristic (ROC) curve was plotted to assess the discriminative capacity of lncRNA DLEU1 and miR-96-5p in identifying DFU. Cell proliferation was detected by CCK-8 assay. Cell apoptosis was assayed by Annexin V-FITC/PI staining method. Bioinformatics, luciferase reporter activity assay, and in vitro cell experiments were used to explore the relationship between lncRNA DLEU1 and miR-96-5p. Results: LncRNA DLEU1 and miR-96-5p were significantly up- and downregulated in patients with DFU, respectively, compared with controls. After ROC assessment, lncRNA DLEU1 and miR-96-5p were found to discriminate DFU from miR-96-5p. Furthermore, lncRNA DLEU1 inhibited human umbilical vein endothelial cells (HUVECs) cell proliferation and increased HUVECs apoptosis and oxidative stress through sponging miR-96-5p. Conclusion: Our findings suggest lncRNA DLEU1 and miR-96-5p as circulating biomarkers for DFU. Also, we provide the clue for the pathogenic significance of lncRNA DLEU1/miR-96-5p in DFU, as well as insights for new potential targets. [ABSTRACT FROM AUTHOR]

Background: C-phycocyanin, a biliprotein from Spirulina platensis, is a future candidate for cancer management. This agent is originated from edible blue-green algae, and numerous in vivo and in vitro research have reported on its anti-cancer properties. The effects of Cphycocyanin have been investigated on caspases 3, 8, 9, and p53 pathways in the human colorectal adenocarcinoma cell line (HT-29) and human umbilical vein endothelial cells (HUVECs). Methods: In the current study, we investigated the effect of C-phycocyanin on caspase 3, 8, 9, and p53-mediated apoptosis pathways in two cell lines (HT-29 & HUVEC), using quantitative real-time PCR and flow cytometry. The cytotoxicity of phycocyanin on HT-29 cells was compared with HUVEC normal cells via colorimetric assays. Results: Based on our findings at molecular level, the expression of caspases 3, 8, 9, and p53 genes were increased in colorectal cancer cells treated with C-phycocyanin. The results were confirmed by an increase in the number of colorectal cancer cells in the early and late stages of apoptosis as compared to the control, untreated cells. In addition, the results of colorimetric assay showed that C-phycocyanin has no cytotoxic effects on normal HUVECs cells. Conclusions: Based on our experimental data, it is evident that C-phycocyanin has measurable effects on cell apoptosis. Since tumorigenesis is halted by apoptosis, C-phycocyanin can be a hopeful candidate for the treatment of human colorectal cancer in the future. [ABSTRACT FROM AUTHOR]

AIM: To investigate the effects of adipose-derived stem cells （ADSCs） with overexpression or silencing of long noncoding RNA（ lncRNA） SNHG8 on the viability, migration, angiogenesis, and the expression of vasoactive factors in human umbilical vein endothelial cells （HUVECs）. METHODS: Identification of ADSCs derived from morbidly obese patients （O-ADSCs） was conducted using flow cytometry and induction of lipogenesis and osteogenesis. The expression of lncRNA SNHG8 in healthy human ADSCs （H-ADSCs） and O-ADSCs was detected by RT-qPCR. Transwell method was used to establish the indirect co-culture system of ADSCs and HUVECs for 48 h, and the cells were divided into O-ADSCs+HUVECs group, H-ADSCs+HUVECs group, and HUVECs alone group. The mRNA and protein expression levels of angiotensin II（ Ang II）, endothelin-1（ ET-1） and endothelial nitric oxide synthase（ eNOS） in HUVECs were detected by RT-qPCR and Western blot. The lncRNA SNHG8 overexpression and silencing lentiviruses were con-structed and used to infect O-ADSCs. The indirect co-cultured ADSCs and HUVECs were divided into O-ADSCs-OE-SNHG8+ HUVECs group, O-ADSCs-OE-NC+HUVECs group, O-ADSCs-sh-SNHG8+HUVECs group, and O-ADSCs-sh-NC+HUVECs group. After co-culture for 48 h, the viability, migration and tubule formation of HUVECs were detected by CCK-8, scratch and angiogenesis assays, respectively. The mRNA and protein expression levels of Ang II, ET-1 and eNOS in HUVECs were detected by RT-qPCR and Western blot, respectively. The nitrate reductase method was used to detect the content of NO in HUVECs. RESULTS:（ 1） The cultured cells were identified as ADSCs.（ 2） Compared with H-ADSCs, lncRNA SNHG8 expression was significantly up-regulated in O-ADSCs（ P<0. 01）.（ 3） Compared with H-ADSCs+HUVECs group and HUVECs group, the mRNA and protein expression levels of Ang II and ET-1 in HUVECs in O-ADSCs+HUVECs group were up-regulated（ P<0. 01）.（ 4） Overexpression of lncRNA SNHG8 in O-ADSCs enhanced the viability, migration and tube formation ability of HUVECs, up-regulated the mRNA and protein expression levels of Ang II and ET-1, down-regulated the mRNA and protein expression levels of eNOS, and decreased the content of NO in HUVECs （P< 0. 05）. However, silencing of lncRNA SNHG8 in O-ADSCs exerted opposite results （P<0. 05）. CONCLUSION: （1） The O-ADSCs can promote endothelial cell viability, migration and tubule formation through paracrine effects.（ 2） The OADSCs with overexpression of lncRNA SNHG8 promote the imbalance of diastolic and contractile factors secreted by endothelial cells, and induce the dysfunction of vascular endothelial cells. [ABSTRACT FROM AUTHOR]

Among the leucocyte subpopulations circulating in peripheral blood of immune‐compromised patients with disseminated Human cytomegalovirus (HCMV) infection, polymorphonuclear leuckocytes (PMNL) and M/M may carry infectious virus. While only in PMNL early HCMV replicative events do occur, monocytes are susceptible to complete virus replication when they enter human organs, where as macrophages become a site of active complete virus replication. In vivo leucocytes and endothelial cells interact continuously, as suggested by several in vitro experimental findings showing the bidirectional HCMV transmission from leucocytes to and from endothelial cells with the critical aid of adhesion molecules. Recently, the neutralising antibody response in sera from subjects with primary HCMV infection was reported to be much higher and earlier than in human embryonic lung fibroblasts (HELF) cells when measured in endothelial cells and epithelial cells, where virus entry is mediated mostly by the pentamer complex gH/gL/pUL128/pUL130/pUL131, whereas it was much lower and delayed when determined in HELF, where virus entry is mediated mostly by the trimer complex gH/gL/gO. Thus, these results suggested that products of UL128L were the molecules primary responsible for the differential neutralising antibody response. This conclusion was confirmed by a series of polyclonal and monoclonal antibodies directed to the components of pUL128L. Very recently, based on two sets of experiments including inhibition and immunoblotting assays, the pentamer complex/trimer complex ratio has been finally identified as the main factor of the neutralising antibody response. This ratio may change with the virus suspension producer and target cell system as well as number of cell culture passages. [ABSTRACT FROM AUTHOR]

Adding dental pulp stem cells (DPSCs) to vascular endothelial cell–formed vessel-like structures can increase the longevity of these vessel networks. DPSCs display pericyte-like cell functions and closely assemble endothelial cells (ECs). However, the mechanisms of DPSC-derived pericyte-like cells in stabilizing the vessel networks are not fully understood. In this study, we investigated the functions of E-DPSCs, which were DPSCs isolated from the direct coculture of human umbilical vein endothelial cells (HUVECs) and DPSCs, and T-DPSCs, which were DPSCs treated by transforming growth factor beta 1 (TGF-β1), in stabilizing blood vessels in vitro and in vivo. A 3-dimensional coculture spheroid sprouting assay was conducted to compare the functions of E-DPSCs and T-DPSCs in vitro. Dental pulp angiogenesis in the severe combined immunodeficiency (SCID) mouse model was used to explore the roles of E-DPSCs and T-DPSCs in vascularization in vivo. The results demonstrated that both E-DPSCs and T-DPSCs possess smooth muscle cell–like cell properties, exhibiting higher expression of the mural cell–specific markers and the suppression of HUVEC sprouting. E-DPSCs and T-DPSCs inhibited HUVEC sprouting by activating TEK tyrosine kinase (Tie2) signaling, upregulating vascular endothelial (VE)–cadherin, and downregulating vascular endothelial growth factor receptor 2 (VEGFR2). In vivo study revealed more perfused and total blood vessels in the HUVEC + E-DPSC group, HUVEC + T-DPSC group, angiopoietin 1 (Ang1) pretreated group, and vascular endothelial protein tyrosine phosphatase (VE-PTP) inhibitor pretreated group, compared to HUVEC + DPSC group. In conclusion, these data indicated that E-DPSCs and T-DPSCs could stabilize the newly formed blood vessels and accelerate their perfusion. The critical regulating pathways are Ang1/Tie2/VE-cadherin and VEGF/VEGFR2 signaling. [ABSTRACT FROM AUTHOR]

AbstractContext Previous studies have highlighted significant therapeutic effects of Qiqilian (QQL) capsule on hypertension in spontaneously hypertensive rats (SHRs); however, its underlying molecular mechanism remains unclear.Obejective We investigated the potential mechanism by which QQL improves hypertension-induced vascular endothelial dysfunction (VED).Materials and methods In vivo, SHRs were divided into four groups (20 per group) and were administered gradient doses of QQL (0, 0.3, 0.6, and 1.2 g/kg) for 8 weeks, while Wistar Kyoto rats were used as normal control. The vascular injury extent, IL-1β and IL-18 levels, NLRP3, ASC and caspase-1 contents were examined. In vitro, the effects of QQL-medicated serum on angiotensin II (AngII)-induced inflammatory and autophagy in human umbilical vein endothelial cells (HUVECs) were assessed.Result Compared with the SHR group, QQL significantly decreased thickness (125.50 to 105.45 μm) and collagen density (8.61 to 3.20%) of arterial vessels, and reduced serum IL-1β (96.25 to 46.13 pg/mL) and IL-18 (345.01 to 162.63 pg/mL) levels. The NLRP3 and ACS expression in arterial vessels were downregulated (0.21- and 0.16-fold, respectively) in the QQL-HD group compared with the SHR group. In vitro, QQL treatment restored NLRP3 and ASC expression, which was downregulated approximately 2-fold compared with that of AngII-induced HUVECs. Furthermore, QQL decreased LC3II and increased p62 contents (p

Pathologic retinal neovascularization is a potentially blinding consequence seen in many common diseases including diabetic retinopathy, retinopathy of prematurity, and retinal vaso-occlusive diseases. This study investigates epithelial membrane protein 2 (EMP2) and its role as a possible modulator of angiogenesis in human retinal pigment epithelium (RPE) under hypoxic conditions. To study its effects, the RPE cell line ARPE-19 was genetically modified to either overexpress EMP2 or knock down its levels, and RNA sequencing and western blot analysis was performed to confirm the changes in expression at the RNA and protein level, respectively. Protein expression was evaluated under both normoxic conditions or hypoxic stress. Capillary tube formation assays with human umbilical vein endothelial cells (HUVEC) were used to evaluate functional responses. EMP2 expression was found to positively correlate with expression of pro-angiogenic factors HIF1α and VEGF at both mRNA and protein levels under hypoxic conditions. Mechanistically, EMP2 stabilized HIF1α expression through downregulation of von Hippel Lindau protein (pVHL). EMP2 mediated changes in ARPE-19 cells were also found to alter the secretion of a paracrine factor(s) in conditioned media that can regulate HUVEC migration and capillary tube formation in in vitro functional angiogenesis assays. This study identifies EMP2 as a potential mediator of angiogenesis in a human RPE cell line. EMP2 levels positively correlate with pro-angiogenic mediators HIF1α and VEGF, and mechanistically, EMP2 regulates HIF1α through downregulation of pVHL. This study supports further investigation of EMP2 as a promising novel target for therapeutic treatment of pathologic neovascularization in the retina.

BackgroundPersistent inflammation following vascular injury drives neointimal hyperplasia (NIH). Specialized lipid mediators (SPM) mediate resolution which attenuates inflammation and downstream NIH. We investigated the effects of a synthetic analogue of resolvin D1 (RvD1) on vascular cells and in a model of rat carotid angioplasty.MethodsHuman venous VSMC and endothelial cells (EC) were employed in migration, cell shape, toxicity, proliferation and p65 nuclear translocation assays. Murine RAW 264.7 cells were utilized to test the effect of pro-resolving compounds on phagocytic activity. A model of rat carotid angioplasty was used to evaluate the effects of 17R/S-benzo-RvD1 (benzo-RvD1) and 17R-RvD1 applied to the adventitia via 25% Pluronic gel. Immunostaining was utilized to examine Ki67 expression and leukocyte recruitment. Morphometric analysis was performed on arteries harvested 14 days after injury.ResultsExposure to benzo-RvD1 attenuated PDGF- stimulated VSMC migration across a range of concentrations (0.1-100 nM), similar to that observed with 17R-RvD1. Pre-treatment with either Benzo-RvD1 or 17R-RvD1 (10, 100nM) attenuated PDGF-BB-induced VSMC cytoskeletal changes to nearly baseline dimensions. Benzo-RvD1 demonstrated modest anti-proliferative activity on VSMC and EC at various concentrations, without significant cytotoxicity. Benzo-RvD1 (10nM) inhibited p65 nuclear translocation in cytokine-stimulated EC by 21% (p

Vascular endothelial cells are exposed to shear stresses with disturbed vs. laminar flow patterns, which lead to proinflammatory vs. antiinflammatory phenotypes, respectively. Effective treatment against endothelial inflammation and the consequent atherogenesis requires the identification of new therapeutic molecules and the development of drugs targeting these molecules. Using Connectivity Map, we have identified vitexin, a natural flavonoid, as a compound that evokes the gene-expression changes caused by pulsatile shear, which mimics laminar flow with a clear direction, vs. oscillatory shear (OS), which mimics disturbed flow without a clear direction. Treatment with vitexin suppressed the endothelial inflammation induced by OS or tumor necrosis factor-α. Administration of vitexin to mice subjected to carotid partial ligation blocked the disturbed flow-induced endothelial inflammation and neointimal formation. In hyperlipidemic mice, treatment with vitexin ameliorated atherosclerosis. Using SuperPred, we predicted that apurinic/apyrimidinic endonuclease1 (APEX1) may directly interact with vitexin, and we experimentally verified their physical interactions. OS induced APEX1 nuclear translocation, which was inhibited by vitexin. OS promoted the binding of acetyltransferase p300 to APEX1, leading to its acetylation and nuclear translocation. Functionally, knocking down APEX1 with siRNA reversed the OS-induced proinflammatory phenotype, suggesting that APEX1 promotes inflammation by orchestrating the NF-κB pathway. Animal experiments with the partial ligation model indicated that overexpression of APEX1 negated the action of vitexin against endothelial inflammation, and that endothelial-specific deletion of APEX1 ameliorated atherogenesis. We thus propose targeting APEX1 with vitexin as a potential therapeutic strategy to alleviate atherosclerosis.

[Figure: see text].

We describe PROPER-seq (protein-protein interaction sequencing) to map protein-protein interactions (PPIs) en masse. PROPER-seq first converts transcriptomes of input cells into RNA-barcoded protein libraries, in which all interacting protein pairs are captured through nucleotide barcode ligation, recorded as chimeric DNA sequences, and decoded at once by sequencing and mapping. We applied PROPER-seq to human embryonic kidney cells, T lymphocytes, and endothelial cells and identified 210,518 human PPIs (collected in the PROPER v.1.0 database). Among these, 1,365 and 2,480 PPIs are supported by published co-immunoprecipitation (coIP) and affinity purification-mass spectrometry (AP-MS) data, 17,638 PPIs are predicted by the prePPI algorithm without previous experimental validation, and 100 PPIs overlap human synthetic lethal gene pairs. In addition, four previously uncharacterized interaction partners with poly(ADP-ribose) polymerase 1 (PARP1) (a critical protein in DNA repair) known as XPO1, MATR3, IPO5, and LEO1 are validated in vivo. PROPER-seq presents a time-effective technology to map PPIs at the transcriptome scale, and PROPER v.1.0 provides a rich resource for studying PPIs.