Endothelial Cell Growth Factor Research Articles

Aberrant expression of proatherogenic cytokines and growth factors in human umbilical vein endothelial cells from newborns of type 2 diabetic women

Objectives:This study reports the levels of cytokines, chemokines, and growth factors previously identified as taking part in the pathology of atherosclerosis in human umbilical vein endothelial cells derived from mothers with type 2 diabetes and compares them with those in human umbilical vein endothelial cells derived from healthy mothers under normal glucose conditions.Methods:Cytokine analysis measures of human umbilical vein endothelial cell lysates were obtained using a multiple analyte profiling (xMAP) assay based on magnetic bead-based technology, using the MAGPIX instrument. The correlation between cytokines, chemokines, and growth factors was examined statistically in human umbilical vein endothelial cells derived from mothers with type 2 diabetes.Results:This study showed that the expression of proinflammatory cytokine interleukin-1 alpha was significantly greater in human umbilical vein endothelial cells derived from mothers with type 2 diabetes than those derived from healthy mothers. The protein level of granulocyte colony-stimulating factor was higher in human umbilical vein endothelial cells derived from mothers with type 2 diabetes than those derived from healthy mothers. A significant positive correlation was demonstrated between the protein expression of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor in human umbilical vein endothelial cells derived from mothers with type 2 diabetes.Conclusion:Diabetes evokes a persistent inflammatory phenotype in human umbilical vein endothelial cells, as indicated by the enhanced production of cytokines and growth factors under normal glucose conditions.

Effect of Selenium nanoparticles in wound healing

Background: Wound healing is a complex process necessary to repairing damaged tissues and preventing infection. Selenium nanoparticles were known by their antioxidant and antimicrobial effect that is important in wound healing. Aim: the aim of this study was to investigate the effect of selenium nanoparticles in reducing and accelerating the wound healing time in mice. Methods: Se NPs were synthesized by a simple wet chemical method. Then the experimental animals were divided into the following groups: Negative control group, Positive control group, selenium nanoparticles control group, wounded mice treated with selenium nanoparticles group. At the end of experiment, mice were scarified& skin tissue samples were collected for biological analysis [Vascular Endothelial cell Growth Factor, Collagenase I and Nitric oxide] and histopathological study. Results: The results showed that the percentage of wound area has been significantly reduced in Se NPs treated group compared to the positive control. The level of Vascular Endothelial Growth Factor and Collagenase I in Se NPs groups were significantly increased compared to positive control while nitric oxide was significantly decreased in Se NPs treated group. Skin tissue showed effect of selenium nanoparticles by regenerating and recavering skin layer. Conclusion: Selenium nanoparticles have an important role in accelerating and reducing wound healing time.

The proatherogenic effect of high salt diet combined with focal hypoperfusion on spontaneous hypertension rat

Objective: To explore the histopathology, monocytes phenotypes and brain mRNA transcription of angiogenic and atherogenic factors preliminarily in spontaneous hypertensive rats (SHRs) fed with high salt diet and subjected to chronic focal hypoperfusion. Methods: A total of 21 SHRs were randomly assigned into SHR with normal diet (SHR-ND group, n=7), SHR fed with high salt (8%) chows (SHR-HSD group, n=14) groups. After induction of high salt diet for 20 weeks, unilateral carotid artery occlusion was applied to one half of SHR-HSD (SHR-HSD-UCAO, n=7) group for 10 weeks to mimic chronic focal cerebral hypoperfusion. The neuropathology, monocytes phenotypes and brain transcription of fibroblast growth factor (FGF-b), platelet-derived endothelial cell growth factor (PD-ECGF), angiogenin (ANG), transforming growth factor-β (TGF-β) and vascular endothelial growth factor A (VEGF-A) among three groups were compared. Results: The systolic blood pressure ((246±12) mmHg vs (220±16) mmHg, P=0.0291, 1 mmHg=0.133 kPa) and diastolic blood pressure ((189±15) mmHg vs (164±12) mmHg, P=0.0143) of SHR-HSD group were elevated significantly compared with those of SHR-ND group. Compared with normotensive Wistar-Kyoto (WKY), SHR-ND, SHR-HSD and SHR-HSD-UCAO groups demonstrated lipohyalinosis, vessel wall thickening, lumen narrowing and multiple enlarged perivascular space and diffuse disarrangement of nerve fiber and myelin vacuolation in corpus callosum pathologically. The ratio of CD11b(+) CD68(+) monocytes in peripheral blood of SHR-HSD group was higher compared with both SHR-ND and SHR-HSD-UCAO groups (P=0.000 8). The mean inflorescence index (MFI) of CD86 and CD206 showd considerable decline in SHR-HSD-UCAO group compared with those of SHR-HSD group (P=0.018 7 and 0.016 8, respectively). The CD86 MFI of CD11b+CD68+ monocytes in SHR-HSD-UCAO group was remarkably higher than that of SHR-ND and SHR-HSD groups (P=0.000 5). Compared with SHR-ND and SHR-HSD groups, the brain mRNA transcription of angiogenic factors including PD-ECGF and ANG were down-regulated (P=0.004 6 and 0.000 2, respectively), while the atherogenic factors including TGF-β and VEGF-A were up-regulated in SHR-HSD-UCAO group (P<0.000 1 and P=0.045, respectively). Conclusion: SHR-HSD-UCAO group shares the pathophysiological characteristics with advanced stage arteriosclerotic cerebral small vessel disease (aCSVD), including neuropathology, imbalanced circulating monocytes phenotypes and down-regulated angiogenic factors.

Correlation between Vascular Endothelial Cell Growth Factor (VEGF) and Muscular Ultrasonography Finding in Knee Osteoarthritis

Background: Osteoarthritis (OA) is a degenerative joint disorder affecting articular cartilage that may be associated with synovial inflammation. The vascular endothelial growth factor is implicated in the pathogenesis of OA. This study aimed to assess man serum vascular endothelial growth factor (VEGF) levels in patients with primary knee osteoarthritis (KOA) and to evaluate their correlation with clinical manifestations, physical function and musculoskeletal-ultrasonography findings. Results: This is a case control study conducted on 50 subjects, thirty patients with primary Knee OA recruited from the outpatients’ clinic of the Rheumatology, Rehabilitation and Physical Medicine Department Benha University Hospitals, from November 2018 to November 2019. Twenty age and sex matched apparently healthy adult individuals were also included as a control group. The present study showed a highly statically significant difference (p < 0.0001) between the mean serum levels of VEGF in the patients’ and the control groups being higher in the patients group compared to the control group (<0.001). The current study revealed a strong significant positive correlations of VEGF levels with the presence of Baker’s cysts, cartilage thickness index and color Doppler US grading (p < 0.0001). Significant positive correlations were also reported between VEGA levels (p=<0.001) and WOMAC total and WOMAC sub scores. Conclusions: VEGF serum levels are highly correlated with clinical, functional and musculoskeletal ultrasound severity of KOA. Thus it could be used as a potential biomarker for estimating Knee OA disease status

Impacts of chitosan oligosaccharide (COS) on angiogenic activities

It has been proved that chitosan oligosaccharide (COS) has a more favorable therapeutic applications such as wound healing and anti-tumor treatment, and can affect angiogenesis. For better understanding the effect of COS on angiogenic activities at cellular level, COS with different concentration and degree of polymerization (DP) were used to culture human umbilical vein endothelial cells (HUVECs) in this work. Cell proliferation activity, cell morphology, cell migration and angiogenesis associated factor expression of HUVECs were evaluated. The results indicated that COS at a high concentration of 400 μg/mL (COS(400)) and DP of 6 (Chitinhexaose Hydrochloride, COS6) had inhibitory effect on angiogenic activities of HUVECs. Specifically, COS(400) and COS6 inhibited cell proliferation activity, cell migration, and vascular endothelial cell growth factor (VEGF) expression of HUVECs. While COS at a low concentration (<400 μg/mL) and suitable polymerization degrees (DP < 6) had little significant effect on cell proliferation, migration, and VEGF expression of HUVECs, showing dose-dependent effect. These findings provided insight for the potential use of COS, for broadening its future applications in biomedical fields and functional materials area. It also helped guide the design and synthesis of chitosan-based materials as an angiogenesis inhibitor for anti-angiogenic therapy.

Choroidal thickening in retinal vein occlusion patients with serous retinal detachment

To evaluate choroid thickness and macular retinal metrics in treatment naive retinal vein occlusion (RVO) patients with serous retinal detachment (SRD) before and after intravitreal anti-vascular endothelial cell growth factor (VEGF) injection and to elucidate the possible role of choroid in the development of SRD and the potential role of SRD as a prognostic parameter. This is a retrospective study involving 85 RVO patients, 41 central retinal vein occlusion (CRVO), and 44 branch retinal vein occlusion (BRVO), with macular edema: 21 central retinal vein occlusion and 22 branch retinal vein occlusion with SRD and the rest without SRD. Patients were evaluated with ophthalmic examinations and swept-source optical coherence tomography (SS-OCT) both before and 4–6 weeks after intravitreal anti-VEGF treatment. Choroid thickness and retinal metrics were measured and compared between SRD and non-SRD groups within each RVO subtype. In both CRVO and BRVO patients, the mean central subfield foveal thickness (CSFT) and central subfoveal choroid thickness (CSCT) of the SRD groups were thicker than those in the non-SRD groups (p < 0.05) at onset. After one anti-VEGF injection, CSFT and FNRT decreased in all groups (p < 0.05). The CSCTs were thicker in the SRD groups compared with the non-SRD groups (p < 0.05). The mean changes of CSFT were more remarkable in the SRD groups (p < 0.05). Thicker choroid was a feature of naive RVO patients with SRD and SRD may be an indicator of better anatomical recovery of retina in RVO patients after a single dose of anti-VEGF treatment.

Dissecting VEGF-Induced Acute Versus Chronic Vascular Hyperpermeability: Essential Roles of Dimethylarginine Dimethylaminohydrolase-1

Abnormal vascular leakiness plays crucial roles in several pathogenesis including cancer, macular degeneration, cardiovascular diseases and even in COVID-19-related pulmonary edema. Vascular endothelial cell growth factor (VEGF), also well-known as vascular permeability factor (VPF), activates VEGF receptor 2 (VEGFR2) and its downstream signaling leading to both acute and chronic vascular permeability. While many key downstream components are identified in cultured cells and murine models upon acute VEGF administration, the key molecular mediators of VEGF-induced chronic permeability remain unclear due to lack of appropriate in experimental model. Here we demonstrated that chronic VEGF exposure induces unique pattern of signaling as compared to that of acute exposure, including phosphorylation of VEGFR2, c-Src, and steady increases of nitric oxide (NO). Utilizing heat-inducible VEGF transgenic zebrafish (Danio rerio) and establishing an algorithm incorporating segmentation techniques to precisely quantify the extravasated dye, we monitored acute and chronic VEGF-induced vascular hyperpermeability in real time. Importantly, knockdown of dimethylarginine dimethylaminohydrolase-1 (DDAH1), an enzyme essential for NO generation, significantly altered downstream signaling pathways due to both acute and chronic VEGF exposure in human endothelial cells and the VEGF-inducible zebrafish permeability model. Finally, the effect of DDAH1 on VEGF-induced hyperpermeability was confirmed using DDAH1 transgenic mice as well as DDAH1 knockout mice by Miles assays, which exhibited increased and decreased leakage of Evan’s blue dye, respectively. Taken together, our data reveal novel divergent signaling of VEGF-induced acute versus chronic hyperpermeability and identify DDAH1 as a critical player and potentially a therapeutic target of vascular hyperpermeability-mediated pathogenesis.

Amelioration of testosterone-induced benign prostatic hyperplasia using febuxostat in rats: The role of VEGF/TGFβ and iNOS/COX-2

Benign prostatic hyperplasia (BPH) is a common male disorder. Febuxostat is a non-purine, selective inhibitor of xanthine oxidase (XO), which has a strong antioxidant capacity and pleiotropic pharmacological properties. This study's objective was to explore the potential ameliorative effects of febuxostat against testosterone-induced BPH in rats. Febuxostat (10 mg/kg/day, per os [p.o.]) prevented increased prostate index levels, serum levels of prostate-specific antigen (PSA), and testosterone levels compared to animals treated with testosterone alone, when administered for 28 days. Histological examination indicated that febuxostat dramatically ameliorated pathological changes in the prostate architecture compared to the testosterone group. Similarly, febuxostat markedly improved testosterone-induced oxidative stress by inhibiting the increase in lipid peroxide and nitrite content, and by reducing the level of depletion of reduced glutathione (GSH) and superoxide dismutase (SOD) activity, which significantly reduced the prostate content of pro-inflammatory cytokines, including tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6). Furthermore, febuxostat significantly reduced the prostatic content, both in terms of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) messenger ribonucleic acid (mRNA) levels, and of protein levels. Moreover, compared to the testosterone group, febuxostat's beneficial effects prevented the increase in growth factors, comprising vascular endothelial cell growth factor A (VEGF-A) and transforming growth factor beta (TGF-β) protein levels. Its ameliorating effects were equal to those of finasteride, which is the most widely used remedy for BPH. In conclusion, this study provides novel evidence that febuxostat experimentally attenuates testosterone-induced BPH in rats, at least in part by inhibiting iNOS/COX-2 and VEGF/TGF-β pathways.

Adjuvant apatinib treatment after resection of hepatocellular carcinoma with portal vein tumor thrombosis: a phase II trial.

BackgroundSurvival after resection of hepatocellular carcinoma (HCC) with portal vein tumor thrombosis (PVTT) still remains poor. Apatinib, a vascular endothelial cell growth factor receptor 2 inhibitor, has been shown to be safe and effective in patients with advanced HCC, so in the present study its efficacy and safety in the adjuvant setting was explored.MethodsIn this single-center, open-label phase II trial, the patients received apatinib (500 mg/day) until they experienced disease recurrence or intolerable toxicity. The primary endpoint was recurrence-free survival (RFS); the secondary endpoints included overall survival (OS) and safety.ResultsFrom a total of 49 patients who were screened between August 2017 and December 2018, 30 study participants received apatinib. According to the Liver Cancer Study Group of Japan classification of PVTT, there were 7, 11, and 12 participants with Vp1, Vp2, and Vp3, respectively. The median duration of treatment was 4.8 months [interquartile range (IQR): 2.0–8.8], and the median dose of apatinib was 339.7 mg/day (IQR: 267.7–500 mg/day). The median follow-up was 14.3 months (IQR: 12.3–19.3). The median RFS was 7.6 months [95% confidence interval (CI): 5.7–9.5 months]. The 1-year RFS rate and the 1-year OS rate were 36.1% and 93.3%, respectively. A total of 29 (96.7%) patients experienced adverse events, and 14 (46.7%) had grade 3 or 4 adverse events. No treatment-related deaths occurred.ConclusionsApatinib was well tolerated in patients after resection of HCC with PVTT. The median RFS in this group was improved compared with that previously reported.Trial registrationNo.: NCT03261791 (ClinicalTrials.gov).

Significant Decreased Expressions of CaN, VEGF, SLC39A6 and SFRP1 in MDA-MB-231 Xenograft Breast Tumor Mice Treated with Moringa oleifera Leaves and Seed Residue (MOLSr) Extracts.

Moringa oleifera is a miracle plant with many nutritional and medicinal properties. Chemopreventive values of the combined mixture of moringa leaves and seed residue (MOLSr) at different ratios (M1S9, M1S1 and M9S1) were investigated. MOLSr extracts were subjected to phytochemical screening, antioxidant assays, metabolite profiling and cytotoxic activity on the primary mammary epithelial cells (PMECs), non-malignant Chang’s liver cells and various human cancer cell lines (including breast, cervical, colon and liver cancer cell lines). The MOLSr ratio with the most potent cytotoxic activity was used in xenograft mice injected with MDA-MB-231 cells for in vivo tumorigenicity study as well as further protein and gene expression studies. M1S9, specifically composed of saponin and amino acid, retained the lowest antioxidant activity but the highest glucosinolate content as compared to other ratios. Cell viability decreased significantly in MCF-7 breast cancer cells and PMECs after treatment with M1S9. Solid tumor from MDA-MB-231 xenograft mice was inhibited by up to 64.5% at third week after treatment with high-dose M1S9. High-dose M1S9 significantly decreased the expression of calcineurin (CaN) and vascular endothelial cell growth factor (VEGF) proteins as well as the secreted frizzled-related protein 1 (SFRP1) and solute carrier family 39 member 6 (SLC39A6) genes. This study provides new scientific evidence for the chemoprevention potential of MOLSr extracts in a breast cancer model; however, the precise mechanism warrants further investigation.

Stepwise differentiation and functional characterization of human induced pluripotent stem cell-derived choroidal endothelial cells

BackgroundEndothelial cells (ECs) are essential regulators of the vasculature, lining arteries, veins, and capillary beds. While all ECs share a number of structural and molecular features, heterogeneity exists depending on their resident tissue. ECs lining the choriocapillaris in the human eye are lost early in the pathogenesis of age-related macular degeneration (AMD), a common and devastating form of vision loss. In order to study the mechanisms leading to choroidal endothelial cell (CEC) loss and to develop reagents for repairing the choroid, a reproducible in vitro model, which closely mimic CECs, is needed. While a number of protocols have been published to direct induced pluripotent stem cells (iPSCs) into ECs, the goal of this study was to develop methods to differentiate iPSCs into ECs resembling those found in the human choriocapillaris specifically.MethodsWe transduced human iPSCs with a CDH5p-GFP-ZEO lentiviral vector and selected for transduced iPSCs using blasticidin. We generated embryoid bodies (EBs) from expanded iPSC colonies and transitioned from mTESR™1 to EC media. One day post-EB formation, we induced mesoderm fate commitment via addition of BMP-4, activin A, and FGF-2. On day 5, EBs were adhered to Matrigel-coated plates in EC media containing vascular endothelial cell growth factor (VEGF) and connective tissue growth factor (CTGF) to promote CEC differentiation. On day 14, we selected for CECs using either zeocin resistance or anti-CD31 MACS beads. We expanded CECs post-selection and performed immunocytochemical analysis of CD31, carbonic anhydrase IV (CA4), and RGCC; tube formation assays; and transmission electron microscopy to access vascular function.ResultsWe report a detailed protocol whereby we direct iPSC differentiation toward mesoderm and utilize CTGF to specify CECs. The CDH5p-GFP-ZEO lentiviral vector facilitated the selection of iPSC-derived ECs that label with antibodies directed against CD31, CA4, and RGCC; form vascular tubes in vitro; and migrate into empty choroidal vessels. CECs selected using either antibiotic selection or CD31 MACS beads showed similar characteristics, thereby making this protocol easily reproducible with or without lentiviral vectors.ConclusionECs generated following this protocol exhibit functional and biochemical characteristics of CECs. This protocol will be useful for developing in vitro models toward understanding the mechanisms of CEC loss early in AMD.

Optical Control of a CRISPR/Cas9 System for Gene Editing by Using Photolabile crRNA.

Currently CRISPR/Cas9 is a widely used efficient tool for gene editing. Precise control over the CRISPR/Cas9 system with high temporal and spatial resolution is essential for studying gene regulation and editing. Here, we synthesized a novel light-controlled crRNA by coupling vitamin E and a photolabile linker at the 5' terminus to inactivate the CRISPR/Cas9 system. The vitamin E modification did not affect ribonucleoprotein (RNP) formation of Cas9/crRNA/tracrRNA complexes but did inhibit the association of RNP with the target DNA. Upon light irradiation, vitamin E-caged crRNA was successfully activated to achieve light-induced genome editing of vascular endothelial cell-growth factor A (VEGFA) in human cells through a T7E1 assay and Sanger sequencing as well as gene knockdown of EGFP expression in EGFP stably expressing cells. This new caging strategy for crRNA could provide new methods for spatiotemporal photoregulation of CRISPR/Cas9-mediated gene editing.

Preconditioning Effect of High-Intensity Interval Training (HIIT) and Berberine Supplementation on the Gene Expression of Angiogenesis Regulators and Caspase-3 Protein in the Rats with Myocardial Ischemia-Reperfusion (IR) Injury.

Objective It has been shown that angiogenesis is a desirable treatment for patients with ischemic heart disease. We set out to investigate the impact of high-intensity interval training (HIIT) and berberine supplementation on the gene expression of angiogenesis-related factors and caspase-3 protein in rats suffering from myocardial ischemic-reperfusion injury. Methods Fifty rats were divided into the following groups: (1) trained, (2) berberine supplemented, (3) combined, and (4) IR. Each cohort underwent five sessions of HIIT per week for a duration of 8 weeks followed by induction of ischemia. Seven days after completion of reperfusion, changes in the gene expression of angiogenesis-related factors and caspase-3 protein were evaluated in the heart tissue. Results We observed a significant difference between four groups in the transcript levels of vascular endothelial cell growth factor (VEGF), fibroblast growth factor-2 (FGF2), and thrombospondin-1(TSP-1) (p ≤ 0.05). However, the difference in endostatin (ENDO) levels was not significant among the groups despite a discernible reduction (p ≥ 0.05). Moreover, caspase-3 protein and infarct size were significantly reduced in the intervention groups (p ≤ 0.05), and cardiac function increased in response to these interventions. Conclusion The treatments exert their effect, likely, by reducing caspase-3 protein and increasing the expression of angiogenesis-promoting factors, concomitant with a reduction in inhibitors of the process.

A Novel Predictive Model of Diabetic Macular Edema Based on Plasma Cytokines via Machine Learning Algorithms

Objective: The study aimed to investigate the change of plasma cytokines and to build a prediction model for diabetic macular edema (DME) in type 2 diabetes mellitus (T2DM). Methods: In pilot cohort, plasma samples were selected from 15 DM patients and 9 DME patients for screening changes of angiogenesis and inflammation related cytokines by the human cytokine antibody array. These screening cytokines were further validated by ELISA in validation cohort, which containing 100 T2DM with DME (DME group) and 100 T2DM without DME (T2DM group) patients. Pearson correlation were calculated to investigate relationship between cytokines and clinical characteristics. Finally, a prediction model based on plasma cytokines was built by machine learning algorithms. Results: In pilot cohort, 12 plasma cytokines were significant higher, among which there was obvious increase of platelet-derived growth factor-BB (PDGF-BB), tissue inhibitors of metalloproteinase 1 (TIMP-1), angiopoietin 1 (ANG-1), CXC-chemokine ligand 16 (CXCL16) and vascular endothelial cell growth factor receptor 2 (VEGF R2) in DME group (Foldchange > 6 and adjusted p < 0.05), comparing with T2DM group. PDGF-BB, TIMP-1, ANG-1, CXCL16 and VEGFR2 were further confirmed significant elevated in validation cohort. Machine learning algorithms employing random forests (RF) yielded the best performance. In the test set, accuracy, sensitivity, specificity, AUROC and f1 score of RF model were 83.3%, 86.2%, 80.6%, 0.83 and 0.83, respectively. Conclusion: Plasma cytokines were closely association with DME. The prediction model consisting of 4 plasma cytokines in the study was a novel tool for predicting DME in T2DM patients. Funding Statement: This study was funded by Science and Technology Committee of Tongzhou District (Number: KJ2019CX014-25). Declaration of Interests: The authors declare they have no conflict of interest. Ethics Approval Statement: In our research, all procedures complied with the Helsinki Declaration for investigation of human subjects. The study received ethical approval from the competent Institutional Review Boards of Beijing Luhe Hospital. All patient-related procedures were performed strictly following the approval of the internal review and ethics boards of the indicated hospitals.

Insight on Ameliorative Role of Selenium Nanoparticles and Niacin in Wound Healing on Adult Female Albino Mice

Background: Wound healing is a complex process necessary for repairing damaged tissues and preventing infection. Selenium nanoparticles (Se NPs) were known due to their antioxidant and antimicrobial effects, also niacin has angiogenesis and antioxidant effects that are important in wound healing. Objective: The present study was conducted to investigate the effect of Se NPs and niacin in reducing and accelerating the wound healing time in mice. Methods: A simple wet chemical method has been modified to synthesize Se NPs in order to investigate their effect and niacin on reducing the wound healing in 80 adult female albino mice (250 mm2 full thickness open excision wound) that were divided into eight groups (10 mice/each). After 30-days, the mice were sacrificed, blood and tissue samples were taken for analysis. Results: The results showed that the percentage of wound area had been significantly reduced in Se NPs and niacin treated groups compared to the positive control. The level of Vascular Endothelial cell Growth Factor and Collagenase I in Se NPs and niacin groups significantly exceed those of other groups while Nitric Oxide (NO) was significantly decreased in treated groups. Liver and kidney functions showed the lower toxicity effect of Se NPs and niacin. Skin tissue showed the wound healing effect of Se NPs and niacin by regenerating skin layer compared to the positive group. Conclusion: Se NPs and niacin play an important role in accelerating and reducing the time of wound healing while they were antagonistic to each other.

Molecular and Mechanical Mechanisms Regulating Ductus Arteriosus Closure in Preterm Infants.

Failure of ductus arteriosus closure after preterm birth is associated with significant morbidities. Ductal closure requires and is regulated by a complex interplay of molecular and mechanical mechanisms with underlying genetic factors. In utero patency of the ductus is maintained by low oxygen tension, high levels of prostaglandins, nitric oxide and carbon monoxide. After birth, ductal closure occurs first by functional closure, followed by anatomical remodeling. High oxygen tension and decreased prostaglandin levels mediated by numerous factors including potassium channels, endothelin-1, isoprostanes lead to the contraction of the ductus. Bradykinin and corticosteroids also induce ductal constriction by attenuating the sensitivity of the ductus to PGE2. Smooth muscle cells of the ductus can sense oxygen through a mitochondrial network by the role of Rho-kinase pathway which ends up with increased intracellular calcium levels and contraction of myosin light chains. Anatomical closure of the ductus is also complex with various mechanisms such as migration and proliferation of smooth muscle cells, extracellular matrix production, endothelial cell proliferation which mediate cushion formation with the interaction of blood cells. Regulation of vessel walls is affected by retinoic acid, TGF-β1, notch signaling, hyaluronan, fibronectin, chondroitin sulfate, elastin, and vascular endothelial cell growth factor (VEGF). Formation of the platelet plug facilitates luminal remodeling by the obstruction of the constricted ductal lumen. Vasa vasorum are more pronounced in the term ductus but are less active in the preterm ductus. More than 100 genes are effective in the prostaglandin pathway or in vascular smooth muscle development and structure may affect the patency of ductus. Hemodynamic changes after birth including fluid load and flow characteristics as well as shear forces within the ductus also stimulate closure. Current pharmacological treatment for the closure of a patent ductus is based on the blockage of the prostaglandin pathway mainly through COX or POX inhibition, albeit with some limitations and side effects. Further research for new agents aiming ductal closure should focus on a clear understanding of vascular biology of the ductus.

Effects of interleukin-6 and tumor necrosis factor-α on the expression of angiogenic and collagenolytic factors in premature and mature adipocytes

ObjectiveThe remodeling of the vascular network and collagen in the extracellular matrix is closely associated with the expansion and dysfunction of adipose tissue. In the present study, we investigated the effects of interleukin (IL)-6 and tumor necrosis factor (TNF)-α on the expression of angiogenic factors, collagen, and collagenase and its endogenous inhibitor in premature and mature adipocytes. MethodsPremature and mature adipocytes were differentiated from 3T3-L1 cells and stimulated with IL-6 or TNF-α to mimic the early and late phases of obesity development. The levels of expression of angiogenic factors, including vascular endothelial cell growth factor a (Vegfa), hepatocyte growth factor (Hgf), angiopoietin (Angpt)1, and Angpt2, as well as type I collagen, matrix metallopeptidase (Mmp) 13, and tissue inhibitor of Mmp (Timp) 1, were determined using real-time reverse transcription polymerase chain reaction or enzyme-linked immunosorbent assay. Human umbilical vein endothelial cells were grown with the culture supernatant of adipocytes stimulated with/without IL-6 or TNF-α, and the formation of tube structures was evaluated. ResultsIL-6 and TNF-α induced the expression of Vegfa, Hgf, and Angpt2 and decreased the expression of Angpt1 in premature adipocytes, whereas, they decreased the expression of Vegfa and Hgf in mature adipocytes. The culture supernatant of IL-6- or TNF-α-stimulated premature adipocytes induced the formation of tube structures. IL-6 and TNF-α had no effects on type I collagen expression in both premature and mature adipocytes but suppressed the expression of Mmp13 and Timp1 in mature and premature adipocytes, respectively. ConclusionThe effects of IL-6 and TNF-α on the expression of angiogenic and collagenolytic factors differed between premature and mature adipocytes. This finding suggests that these inflammatory cytokines induce expansion and dysfunction of adipose tissue via angiogenesis and collagen turnover in premature and mature adipocytes.

Alkaline phosphatase dual-binding sites for collagen dictate cell migration and microvessel assembly in vitro.

Interactions between cell types, growth factors, and extracellular matrix components involved in angiogenesis are crucial for new vessel formation leading to tissue regeneration. This study investigated whether cocultures of fibroblasts and endothelial cells (ECs; from macro- or microvasculature) play a role in the formation of microvessel-like structures by ECs, as well as modulate fibroblast differentiation and growth factors production (vascular endothelial cell growth factor, basic fibroblast growth factor, active transforming growth factor-β1, and interleukin-8), which are important for vessel sprouting and maturation. Data obtained revealed that in vitro coculture systems of fibroblasts and human ECs stimulate collagen synthesis and growth factors production by fibroblasts that ultimately affect the formation and distribution of microvessel-like structures in cell cultures. In this study, areas with activated fibroblasts and high alkaline phosphatase (ALP) activity were also observed in cocultures. Molecular docking assays revealed that ALP has two binding positions for collagen, suggesting its impact in collagen proteins' aggregation, cell migration, and microvessel assembly. These findings indicate that bioinformatics and coculture systems are complementary tools for investigating the participation of proteins, like collagen and ALP in angiogenesis.

Circular RNA COL1A2 promotes angiogenesis via regulating miR-29b/VEGF axis in diabetic retinopathy

AimsThe dysregulation of circular RNAs (circRNAs) has been implicated in the progression of diabetic retinopathy (DR). This study aims to explore the role and underlying mechanism of hsa_circ_0081108 (circCOL1A2) in DR. Materials and methodscircCOL1A2, vascular endothelial growth factor (VEGF) and miR-29b expression levels in human retinal microvascular endothelial cells (hRMECs) were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blotting. The biological functions of hRMECs were evaluated by MTT, transwell, tube formation, and vascular permeability assays, respectively. The interaction between miR-29b and circCOL1A2/VEGF was determined by dual luciferase assay. The release of VEGF was examined by ELISA. The in vivo role of circCOL1A2 was further verified in streptozotocin (STZ)-induced DR in mice. The pathological changes and VEGF expression in retinal tissues were detected by hematoxylin and eosin (HE) and immunohistochemical staining. Key findingsHigh glucose (HG) challenge led to increased circCOL1A2, VEGF, MMP-2, MMP-9 levels, but decreased miR-29b level in hRMECs. In addition, circCOL1A2 sponged miR-29b to promote VEGF expression. Silencing of circCOL1A2 inhibited HG-induced proliferation, migration, angiogenesis and vascular permeability of hRMECs via enhancing miR-29b expression. Moreover, circCOL1A2/miR-29b axis participated in HG-induced increase in angiogenesis-related protein expression. Finally, circCOL1A2 knockdown suppressed angiogenesis via regulating miR-29b/VEGF axis in DR mice. SignificancecircCOL1A2 facilities angiogenesis during the pathological progression of DR via regulating miR-29b/VEGF axis, suggesting that targeting circCOL1A2 may be a potential treatment for DR.

Protective effect of Shenfu injection on the endothelium of severe sepsis by inhibiting CD11b+ cell paralysis induced by high mobility group protein B1

To investigate the relationship between the biochemical parameters, the pulmonary pathologic injury and the immune mechanism of severe sepsis in infant porcine, and the intervention effect of Shenfu injection. Panamanian infant porcine (2-3 months old) were divided into sham operation group (Sham group; intravenous injection of normal saline), lipopolysaccharide (LPS) induced severe sepsis model group (LPS group; intravenous injection of LPS 1 mg/kg, and continuing at 0.5 mg×kg-1×h-1 for 12 hours), and Shenfu injection intervention group (SF group; intravenous injection of Shenfu injection 10 mL/kg at the same time of modeling, twice a day) according to the random number table method, with 5 in each group. Forty-eight hours after the challenge, the changes of C-reactive protein (CRP), procalcitonin (PCT), oxygenation index (PaO2/FiO2), base excess (BE), blood lactate (Lac) and other biochemical indexes were detected with blood sampling; the number of neutrophils [myeloperoxidase positive (MPO+)] and their activated subsets CD11b+CD64+, M1 macrophages (CD80+CD64+), CD4+ and CD8+ T cells were analyzed in peripheral blood by flow cytometry. The levels of plasma cytokines were detected by enzyme linked immunosorbent assay (ELISA). The pathological damage of lung tissue was observed by hematoxylin-eosin (HE) staining. The mRNA expression of lung injury related molecules and their receptors, chemokines, cytokines, vascular endothelial related molecules and tight junction protein were detected by reverse transcription-polymerase chain reaction (RT-PCR). Compared with Sham group, the levels of CRP, PCT and Lac in LPS group significantly increased, and PaO2/FiO2 and BE significantly decreased. In the peripheral blood, CD80+CD64+ macrophages, CD11b+CD64+ and MPO+ neutrophils, CD4+, CD8+ T cells and tumor necrosis factor-α (TNF-α) significantly decreased, but interleukin-10 (IL-10) and vascular endothelial cell growth factor (VEGF) significantly increased; the mRNA expressions of high mobility group protein B1 (HMGB1), receptor for advanced glycation end products (RAGE), angiopoietin 2/angiopoietin 1 (Ang2/Ang1) significantly increased, Toll like receptor 9 (TLR9), chemokines (CXCL9 and CXCL10), TNF-α, IL-27, Tek tyrosine kinase 2 (TIE2), vascular endothelial cadherin (VE-CAD) and Occludin significantly decreased in lung tissue. HE staining showed inflammatory cell infiltration and exudation in the alveolar cavity, alveoli consolidation, thickening of the alveolar interstitial layer and emphysema. Compared with LPS group, Lac in SF group significantly decreased (mmol/L: 4.2±1.0 vs. 6.3±1.1, P < 0.05), while BE and PaO2/FiO2 significantly increased [BE (mmol/L): -6.4±2.6 vs. -11.6±2.5, PaO2/FiO2 (mmHg, 1 mmHg = 0.133 kPa): 180±36 vs. 105±35, both P < 0.05]; the percentage of CD80+CD64+ macrophages, CD11b+CD64+ and MPO+ neutrophils significantly increased [CD80+CD64+: (7.13±2.01)% vs. (3.80±0.46)%, CD11b+CD64+: (8.33±2.55)% vs. (2.15±0.47)%, MPO+: (21.22±2.33)% vs. (8.31±0.46)%, all P < 0.05]; the mRNA expressions of HMGB1 and RAGE decreased to some extent [HMGB1 mRNA (2-ΔΔCT): 1.81±0.45 vs. 2.23±0.85, RAGE mRNA (2-ΔΔCT): 6.69±3.48 vs. 11.60±6.91, both P < 0.05], the mRNA expressions of CXCL9 and TIE2 increased to a certain extent [CXCL9 mRNA (2-ΔΔCT): 1.06±0.63 vs. 0.50±0.12, TIE2 mRNA (2-ΔΔCT): 1.42±0.68 vs. 0.27±0.16, both P < 0.05]; the pathological damage of lung tissue were significantly alleviated. The increased abnormality of BE, Lac, and PaO2/FiO2, the immune exhausting and paralysis may be important factors leading to the fatal lung injury in infant porcine with severe sepsis. The possible mechanism is that the excessive activation of HMGB1 and its receptor RAGE, the suppression of TLR9 and corresponding chemokine and inflammatory factors lead to increased endothelial damage and decreased tight connection, which in turn induces capillary leakage. The intervention of immune disorders by Shenfu injection in the severe pneumonia accompanied by sepsis may be related to elevated M1 macrophages and activated neutrophils in the peripheral blood, inhibition of HMGB1 and its receptor RAGE mRNA expression, and elevated CXCL9 and TIE2 expression.