Vascular Endothelial Growth Factor Expression In Cells Research Articles

Inhibition of PDGFRβ alleviates endothelial cell apoptotic injury caused by DRP-1 overexpression and mitochondria fusion failure after mitophagy

Kawasaki disease (KD), described as “mucocutaneous lymph node syndrome”, affects infants and toddlers. Patients with KD suffer from an inflammatory cascade leading to vasculitis with a predilection for coronary arteries. While the symptoms and pathogenesis of KD have received more and more attention, the precise mechanisms are still debated. Researches show that endothelial dysfunction process in KD leads to arterial damage and affect clinical outcome. In this study, we constructed a Candida albicans water soluble fraction (CAWS)-induced KD murine model and penetrated investigating the mechanisms behind endothelial dysfunction. CAWS-induced mice presented remarkably elevated vascular endothelial cell growth factor (VEGF) levels. Abundant expression of VEGF was documented in all vessels that showed edema from acute KD. It has been reported that Platelet-derived growth factor (PDGF) co-expression normalizes VEGF-induced aberrant angiogenesis. Hyperexpression of PDGFRβ was induced in the thickened medial layer and vascular endothelium of KD mice. Masitinib (Mas) is an oral tyrosine kinase inhibitor of numerous targets, which can selectively target PDGFR signaling. We set out to explore whether Mas could regulate coronary pathology in KD. Mas administration significantly reduced the VEGF-induced endothelial cells migration. NOX4 was activated in vascular endothelial cells to produce more ROS. Mitochondrial dysregulated fission and mitophagy caused by DRP-1 overexpression precipitated the arterial endothelial cells injury. Here, mitophagy seemed to work as the driving force of DRP-1/Bak/BNIP3-dependent endothelial cells apoptosis. In summary, how mitophagy is regulated by DRP-1 under pathologic status is critical and complex, which may contribute to the development of specific therapeutic interventions in cardiovascular diseases patients, for example Masatinib, the inhibitor of PDGFRβ.Facts and questionsKawasaki disease causing systemic vasculitis, affects infants and toddlers.Coronary artery injury remains the major causes of morbidity and mortality.DRP-1 overexpression induces DRP-1/Bak/BNIP3-dependent endothelial cells apoptosis.PDGFRβ was high-expressed in the thickened medial layer of CAWS-induced KD mice.Inhibition of PDGFRβ signaling alleviates arterial endothelial cells injury.

The effect of autologous plasminogen on the rate of healing of chronic skin ulcers in patients with diabetes mellitus and the level of proteins - markers of hypoxia and angiogenesis

chronic diabetic wounds are a complication of diabetes mellitus that develop as a result of the toxic effects of hyperglycemia, which leads to metabolic disorders with the development of immunosuppression, angiopathy, and neuropathy. A considerable amount of experimental data has been accumulated, which indicates that the participation of the plasminogen-plasmin system in wound healing is not only to clear wounds of fibrin clots, but also to fine-tune the activity of cells that ensure an adequate course of all stages of healing. This study was conducted to investigate the effect of autologous plasminogen on the rate of chronic wound healing in patients with diabetic foot syndrome. The study involved 45 patients with purulent necrotic form of diabetic foot. The efficacy of topical application of standard medications was compared with the application of autologous plasminogen. The wound area was measured once every two days until complete healing. To study the molecular mechanisms of plasminogen effect on wound healing, the level of marker proteins of key physiological processes associated with healing, namely hypoxia and angiogenesis, was determined. The levels of protein regulators of angiogenesis (vascular endothelial cell growth factor (VEGF), hypoxia-inducible factor-1α (HIF-1α) and angiostatins) in the skin tissue of wounds at the beginning and end of treatment with autologous plasminogen in patients with chronic diabetic wounds with neuropathic diabetic foot were evaluated. The levels of VEGF, HIF-1α and angiostatins were determined by Western blotting. We have found that wound tissue in patients with chronic diabetic wounds before treatment is characterized by a reduced level of VEGF, which represents the inability of these wounds to regenerate. At the same time, an increased level of HIF-1α and angiostatins is detected. After treatment with autologous plasminogen applications, a significant increase in VEGF expression was observed against the background of a progressive decrease in HIF-1α and angiostatin activity. The results indicate that increased production of angiogenic inhibitors may counteract angiogenesis and contribute to the failure of chronic wound healing. The use of plasminogen applications reduces the level of angiostatins in wound biopsies, which allowed to increase the level of proangiogenic factors. Treatment with plasminogen demonstrated an improvement in the condition of the wound surface and the transition of healing to the proliferative phase due to the initiation of the inflammatory process in wounds. The results of planimetric measurement of the wound area indicate a 5-fold reduction in the time required for complete wound healing in the group receiving plasminogen applications compared to this parameter in the comparison group.

Preparation of total triterpenoids from Antrodia cinnamomea fermentation mycelium and their in vitro inhibitory effects on hepatocellular carcinoma

Total triterpenoids from Antrodia cinnamomea fermentation mycelium (TT-ACFM) were prepared, and their in vitro inhibitory effects on hepatocellular carcinoma were investigated. Human hepatocellular carcinoma HepG2 cells were incubated with TT-ACFM with concentration of 0 (control), 12.5, 25, 50, 100 and 200 μg/mL, respectively. The proliferation of cell was detected using CCK-8 method. The apoptosis of cells was detected by flow cytometry. The migration and invasion of cells was determined using Transwell chamber. The level of reactive oxygen species (ROS) in cells was determined using DCFH-DA method. The expressions of matrix metalloproteinse-2 (MMP-2), matrix metalloproteinse-9 (MMP-9) and vascular endothelial growth factor (VEGF) protein in cells were detected by western blot assays. Results indicate that, TT-ACFM can not only inhibit the proliferation of HepG2 cells and promote their apoptosis, but also inhibit their invasion and invasion. The mechanism may be related to its increase of ROS and down-regulation of MMP-2, MMP-9 and VEGF expression in cells.

Fibrin glue: an anti-fibrotic agent for contracted socket

Background: The contracted socket is caused by fibrosis leading to the shortening of the orbital tissue. Vascular endothelial growth factor (VEGF) is a profibrotic factor in postoperative wound healing. The novel option to prevent fibrosis is the use of fibrin glue. This study aims to evaluate the role of fibrin glue in reducing the VEGF expression in cells that cause post-socket surgery scar. Methods: An in vivo experimental study using an animal model was conducted. We used twenty eyes of white rabbit species as the study sample. The evisceration was carried out and distributed to the animals in four experimental groups consisting of the control group, the MMC group (0.1 ml of mitomycin-C (MMC) 0.4 mg/ml), the triamcinolone acetonide (TCA) group (0.1 ml of TCA 40 mg/ml), and the fibrin glue group (0,1 ml of fibrin glue). The treatment was performed through injection of the subconjunctival. After 14 days, rabbits were euthanized, and conjunctiva samples were cut. Immunoreactivity scores (IRS) were used to analyze the expression of VEGF in epithelia conjunctiva with a significant level of p <0.05. Data were analyzed using SPSS version 26 for Windows. Results: All groups showed a statistically significant difference in VEGF expression with a p-value 0,000 (p<0,05). The VEGF level was significantly lower in the three treatment groups compared with the control group. The VEGF expression was highest for the control group, followed by fibrin glue, TCA and MMC group. We found significant differences between the control group and the three treatment groups; the MMC group (p=0.001), the TCA group (p=0.002), and the fibrin glue group (p=0.005). While differences in VEGF expression between the three treatment groups could not be proven statistically. Conclusion: Fibrin glue is proven to reduce VEGF expression as a marker of fibrosis; thus, fibrin glue can be an alternative anti-fibrotic agent.

Role of Vascular Endothelial Cell Growth Factor on Pathophysiology of Uterine Adenomyosis

Background: Uterine adenomyosis is defined as the presence of ectopic endometrial tissue in the myometrium of the uterus and is a known cause of chronic pelvic pain, heavy menstrual bleeding, and subfertility. However, its pathogenesis is not completely established. Several reports have suggested that vascular endothelial cell growth factor (VEGF) may be associated with the progression of adenomyosis. The goal of this study was to evaluate the role of VEGF on pathophysiology of uterine adenomyosis by comparing expression of VEGF in the same uterus and in the endometrium and myometrium, with patients’ adenomyosis. Methods: We analyzed 22 premenopausal patients with a focal type of uterine adenomyosis who received an adenomyomectomy between December 2019 and April 2020 at our hospital. All patients were preoperatively treated with gonadotropin-releasing hormone(GnRH) analogs. During these surgeries, samples were obtained from the uterus of each patient which included the adenomyosis lesion, the myometrium without adenomyosis, and endometrial tissue. Immunohistochemistry stain of VEGF and real-time polymerase chain reaction (RT-PCR) of VEGF expression were compared for each of three points in the uterus. We also compared microvascular density in the adenomyosis lesion between the ectopic endometrial gland and myometrial stroma. Results: VEGF expression was found to be increased in adenomyotic lesions and myometrium compared with the eutopic endometrium (p < 0.0001). In addition, RT-PCR indicated higher VEGF expression in the myometrial and adenomyosis tissues than in the eutopic endometrium (p < 0.05). In each patient, a comparison of microvascular density (MVD) measurements of the eutopic endometrium with an endometrial component and stroma in the adenomyosis tissue indicated a significant increase in the stroma comprising the myometrium (p = 0.02). Conclusions: VEGF contributes to the progression of uterine adenomyosis and may be more strongly activated in the stromal component of the myometrium than in the endometrial (eutopic or ectopic) component of the adenomyosis in the same uterus. This suggests that VEGF plays a significant role in the muscular component of the endometrium during the progression of adenomyosis.

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.

Effect of Platelet-Activating Factor on Barrier Function of ARPE-19 Cells.

AimTo examine the effects of platelet-activating factor (PAF) on the barrier functions of cultured retinal pigment epithelial (RPE) cells.MethodsA human RPE cell line (ARPE-19) was cultured on microporous filter supports and treated with PAF and WEB 2086, a specific PAF-receptor (PAF-R) antagonist. The permeability of the RPE monolayer was measured using transepithelial electrical resistance (TER) and sodium fluorescein flux. The expression of the tight junction protein zonula occludens (ZO)-1 and the adherens junction protein N-cadherin was assessed using immunohistochemistry and Western blotting. We also measured the vascular endothelial growth factor (VEGF) concentrations in PAF-treated cultures and re-measured RPE monolayer permeability in the presence of VEGF-neutralizing antibodies.ResultsPAF significantly decreased the TER and enhanced the sodium fluorescein flux of the RPE monolayer and downregulated the expression of ZO-1 and N-cadherin. These effects were abolished by WEB 2086-mediated blockage of the PAF-R. PAF stimulation increased VEGF expression in RPE cells, and the antibody-mediated neutralization of VEGF caused a partial recovery of the barrier properties.ConclusionThe barrier functions of ARPE-19 cells were altered by PAF, and these effects were partly mediated by an upregulation of VEGF expression in these cells. Our results contribute to the growing body of evidence supporting the role of PAF in choroidal neovascularization. Our findings suggest that PAF is a novel target in the development of therapies for increased permeability of the RPE monolayer.

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.

Chronological changes in rat heel skin following depressurization of pressure ulcer-like dermal lesions

In our previous study, we proposed an animal model in which pressure ulcer-like dermal lesions can be produced by denervation of the sciatic nerve and application of a pressure load to rat heel skin. In the present study, we divided these animals into non-treated and pressure loading groups, and initiated hindlimb unloading (depressurization) by tail suspension at 1, 3, 5, 7, and 14 days after inflicting lesions (1–14d pressurization groups). Chronological changes in heel lesions were examined morphologically in all treatment groups after 1, 3, 7, 14, 28, and 40 days. Open dermal lesions were formed by 14 days in the loading group and numerous macrophages were present. In the 14d pressurization group, numerous macrophages were still distributed in and around lesions and Vascular endothelial cell growth factor (VEGF) expression was strongly detected by 3 days, but a thin germinal layer began to appear and CD68-positive macrophages and VEGF immunoreactions decreased gradually by 7 days later. By 14 days after depressurization, the germinal layer was repaired, and macrophages and immunoreactions of VEGF were similar to those of non-treated skin. These chronological changes were similar to those in human pressure ulcers, but from 5d after depressurization, different chronological changes were observed. Specifically, epidermis was thickened and macrophages were hardly detected at 5 days in the loading group, but the epidermis disappeared by 1 day in the 5d pressurization group. Subsequently, numerous macrophages aggregated and VEGF expression was increased by 3 days, and the remaining healing process was similar to that in the 14d pressurization group. Even when unloading was performed during the early stages (5d pressurization group), the epidermis disappeared and macrophages were then distributed before repair of the lesion was observed. These results suggest that earlier migration of macrophages to skin lesions might be associated with rapid wound healing.

Cooperation of Indian Hedgehog and Vascular Endothelial Growth Factor in Tumor Angiogenesis and Growth in Human Hepatocellular Carcinomas, an Immunohistochemical Study.

The Hedgehog pathway was recently shown to be involved in vascular development and neovascularization in human embryogenesis and disease. However, the role of Hedgehog pathway in modulating tumor angiogenesis is still unexplored. In the current study, we investigated the expression of Indian Hedgehog (Ihh) and vascular endothelial cell growth factor (VEGF) in human hepatocellular carcinomas (HCCs) with immunohistochemical staining and compared the immunoreaction data with various clinicopathologic characteristics. Immunoreactivity of Ihh and VEGF proteins was observed in 61.5% (56/91) and 64.5% (59/91) cases of HCC tumor tissues, respectively, which was considerably higher than the adjacent nonmalignant tissues. Ihh protein was observed predominantly in the cytoplasm of the tumor cells with a staining pattern of which was sparse and dot-like, or circular around the cell membrane. VEGF protein was expressed heterogenously in the cytoplasm in tumor cells and was negative in peritumoral areas in all cases. CD34 showed diffuse staining in the tumor parenchyma in most HCC specimens. The association of expression of Ihh and VEGF with tumor size was statistically significant (P<0.05), but there was no significant association with other clinicopathologic parameters. Moreover, there was a significant association of the expressions of Ihh and VEGF proteins in HCC (r=0.6, P<0.0001), and of Ihh and CD34 staining (r=0.261, P=0.012). Our findings suggest that Ihh is involved in the development of HCC. These findings are also consistent with the concept that cooperation of Ihh and VEGF modulate HCC tumor angiogenesis and growth.

Long-chain noncoding RNA GAS5 mediates oxidative stress in cardiac microvascular endothelial cells injury.

This study is performed to figure out the role of long-chain noncoding RNA growth-arrest specific transcript 5 (GAS5) in homocysteine (HCY)-induced cardiac microvascular endothelial cells (CMECs) injury. CMECs were cultured and the model of CMECs injury was established by coincubation with HCY. To construct stable overexpression of GAS5 cells, the expression of GAS5, microRNA-33a-5p (miR-33a-5p) and ATP-binding cassette transporter A1 (ABCA1), and biological characteristics of cells were determined. The messenger RNA (mRNA) level and secretion of vascular endothelial growth factor (VEGF), activity of reactive oxygen species (ROS) and superoxide dismutase (SOD), and the content of malondialdehyde (MDA) were measured. The binding site between GAS5 and miR-33a-5p and between miR-33a-5p and ABCA1 was verified. CMECs were successfully cultured. Reduction of GAS5 expression and ABCA1 expression together with increased expression of miR-33a-5p was found in CMECs induced by HCY. After overexpression of GAS5, there showed increased proliferative activity, decreased cell apoptosis rate and apoptosis index, enhanced cell migration ability, increased number of lumen formation, increased mRNA expression of VEGF in cells and the secretion in the supernatant, decreased activity of ROS and SOD in cells, and decreased content of ROS in cells. miR-33a-5p could promote the enrichment of GAS5 and ABCA1 was the direct target gene of miR-33a-5p. Our study suggests that the low expression of GAS5 was observed in HCY-induced CMECs injury, and the upregulation of GAS5 could attenuate HCY-induced CMECs injury by mediating oxidative stress, and its mechanism is related to the upregulation of ABCA1 expression by competitively binding with miR-33a-5p.

Protective effect of Danhong Injection combined with Naoxintong Capsule on cerebral ischemia-reperfusion injury in rats

Ethnopharmacological relevanceDanhong Injection (DHI) and Naoxintong Capsule (NXT) are renowned traditional Chinese medicine in China. The drug combination of DHI and NXT is frequently applied for the treatment of cardiovascular and cerebrovascular diseases in clinic. However, there had been no pharmacological experiment studies of interaction between DHI and NXT. Due to the drug interactions, exploring their interaction profile is of great importance. Material and methodsIn this study, focal cerebral I/R injury in adult male Sprague-Dawley rats were induced by transient middle cerebral artery occlusion (tMCAO) for 1h followed by reperfusion. Rats were divided into 5 groups: sham group, ischemia reperfusion untreated group (IRU), DHI group (DHI 10mL/kg/d), NXT group (NXT 0.5g/kg/d), DHI plus NXT group (DHI-NXT, DHI 10mL/kg/d plus NXT 0.5g/kg/d). All drug-treated groups were respectively successive administrated for 7 days after ischemia/ reperfusion (I/R) injury. The effects on rat neurological function were estimated by neurological defect scores. Brain infarct volumes were determined based on 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. Pathological changes in brain tissues were observed using hematoxylin and eosin (H&E) staining and transmission electron microscope (TEM). Levels of nitric oxide (NO), granulocyte colony-stimulating factor (G-CSF) and granulocyte macrophage colony-stimulating factor (GM-CSF) in serum were determined with enzyme-linked immunosorbent assay (ELISA). Immunohisto-chemistry and Western blot were used to detect the expressions of basic fibroblast growth factor (bFGF), von Willebrand factor-microvessel vascular density (vWF-MVD), vascular endothelial cell growth factor (VEGF), transforming growth factor-β1 (TGF-β1), angiogenin-1 (Ang-1), angiogenin-2 (Ang-2) and platelet derived growth factor (PDGF) at day 7 after ischemia/reperfusion (I/R) injury. ResultsCompared with IRU group and mono-therapy group (DHI group or NXT group), Danhong Injection combined with Naoxintong Capsule (DHI-NXT) group significantly ameliorated neurological deficits scores, infarct volume and pathological change, significantly decreased the overexpression of NO and the level of Ang-1, significantly increased the expressions of VEGF, Ang-2, G-CSF, GM-CSF, bFGF, PDGF, vWF, TGF-β1. ConclusionThe protective benefits on rat brain against I/R injury were clearly produced when DHI and NXT were used in combination, which provided rational guidance for clinical combined application of DHI and NXT, and this protection maybe associated with the up-regulation expressions of the related chemokines and growth factors of angiogenesis.

Human umbilical cord mesenchymal stem cells improve the reserve function of perimenopausal ovary via a paracrine mechanism

BackgroundHuman umbilical cord mesenchymal stem cells (hUCMSCs) are a type of pluripotent stem cell which are isolated from the umbilical cord of newborns. hUCMSCs have great therapeutic potential. We designed this experimental study in order to investigate whether the transplantation of hUCMSCs can improve the ovarian reserve function of perimenopausal rats and delay ovarian senescence.MethodWe selected naturally aging rats confirmed by vaginal smears as models of perimenopausal rats, divided into the control group and the treatment group, and selected young fertile female rats as normal controls. hUCMSCs were transplanted into rats of the treatment group through tail veins. Enzyme-linked immunosorbent assay (ELISA) detected serum levels of sex hormones, H&E staining showed ovarian tissue structure and allowed follicle counting, immunohistochemistry and western blot analysis revealed ovarian expression of hepatocyte growth factor (HGF), vascular endothelial cell growth factor (VEGF), and insulin-like growth factor-1 (IGF-1), polymerase chain reaction (PCR) and western blot analysis revealed hUCMSCs expression of HGF, VEGF, and IGF-1.ResultsAt time points of 14, 21, and 28 days after hUCMSCs transplantation, estradiol (E2) and anti-Müllerian hormone (AMH) increased while follicle-stimulating hormone (FSH) decreased; ovarian structure improved and follicle number increased; ovarian expression of HGF, VEGF, and IGF-1 protein elevated significantly. Meanwhile, PCR and western blot analysis indicated hUCMSCs have the capacity of secreting HGF, VEGF, and IGF-1 cytokines.ConclusionsOur results suggest that hUCMSCs can promote ovarian expression of HGF, VEGF, and IGF-1 through secreting those cytokines, resulting in improving ovarian reserve function and withstanding ovarian senescence.

Albendazole inhibits HIF-1α-dependent glycolysis and VEGF expression in non-small cell lung cancer cells.

Albendazole (ABZ) has an anti-tumor ability and inhibits HIF-1α activity. HIF-1α is associated with glycolysis and vascular endothelial cell growth factor (VEGF) expression, which plays an important role in cancer progression. These clues indicate that ABZ exerts an anti-cancer effect by regulating glycolysis and VEGF expression. The aim of this study is to clarify the effects of ABZ on non-small cell lung cancer (NSCLC) cells and explore the underlying molecular mechanisms. The expression levels of HIF-1α and VEGF were detected using western blot analysis, and the effect of ABZ on glycolysis was evaluated by measuring the relative activities of hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) and detecting the production of lactate in A549 and H1299 cells. The results showed that ABZ decreased the expression levels of HIF-1α and VEGF and suppressed glycolysis in under hypoxia, but not normoxic condition. Inhibiting HIF-1α also suppressed glycolysis and VEGF expression. Additionally, ABZ inhibited the volume and weight, decreased the relative activities of HK, PK, and LDH, and reduced the levels of HIF-1α and VEGF of A549 xenografts in mouse models. In conclusion, ABZ inhibited growth of NSCLC cells by suppressing HIF-1α-dependent glycolysis and VEGF expression.

IL-17 Promotes Angiogenic Factors IL-6, IL-8, and Vegf Production via Stat1 in Lung Adenocarcinoma.

Inflammation and angiogenesis are two hallmarks of carcinoma. The proinflammatory cytokine interleukin-17 (IL-17) facilitates angiogenesis in lung cancer; however, the underlying mechanism is not fully understood. In this study, tumour microvessel density (MVD) was positively associated with IL-17, interleukin-6 (IL-6), interleukin-8 (IL-8), and vascular endothelial cell growth factor (VEGF) expression in human lung adenocarcinoma tissues, and it was increased in tumour tissues of A549-IL-17 cell-bearing nude mice. Importantly, positive correlations were also detected between IL-17 expression and IL-6, IL-8 and VEGF expression in human lung adenocarcinoma tissues. Furthermore, IL-6, IL-8 and VEGF production, as well as STAT1 phosphorylation, were increased in tumour tissues of A549-IL-17 cell-bearing nude mice in vivo and in A549 and H292 cells following IL-17 stimulation in vitro. In addition, STAT1 knockdown using an inhibitor and siRNA attenuated the IL-17-mediated increases in IL-6, IL-8 and VEGF expression in A549 and H292 cells. In conclusion, IL-17 may promote the production of the angiogenic inducers IL-6, IL-8 and VEGF via STAT1 signalling in lung adenocarcinoma.

Expression of Survivin and vascular endothelial cell growth factor in transitional cell carcinoma of bladder and its clinical significance

Objective To investigate the expression and clinical significance of Survivin and vascular endothelial cell growth factor (VEGF) in transitional cell carcinoma of bladder (BTCC). Methods The immunohistochemical PV-6000 method was used to study the expression level of Survivin and VEGF protein in 146 cases of BTCC and 23 cases of normal bladder tissues. Results The positive expression rate of Survivin and VEGF protein in BTCC was 69.9% (102/146) and 74.7% (109/146) respectively, which was significantly higher than that of bladder normal tissues (P<0.05). The positive expression of Survivin and VEGF had a positive correlation and both were closely related to the histological grade and clinical stages of BTCC (P<0.05). Conclusion Survivin and VEGF may play an important role in the carcinogenesis and progression of BTCC. Key words: Survivin; Vascular endothelial cell growth factor; Transitional cell carcinoma of bladder; Immunohistochemistry

Mechanism of Wnt/β-catenin signaling pathway in enhanced malignant phenotype of non-small cell lung cancer induced by anti-angiogenesis therapy

To study the mechanism of Wnt/β-catenin signaling pathway in the enhanced malignant phenotype of A549 cells of human non-small cell lung cancer induced by the anti-angiogenesis therapy. The siRNA technique was employed to inhibit the expression of vascular endothelial growth factor (VEGF) in A549 cells and simulate the clinical course of anti-angiogenesis therapy. Real-time PCR and western blot were used to study the change in the expression of Wnt/β-catenin signaling molecules at the mRNA and protein level respectively, as well as the effect on the epithelial mesenchymal transition in A549 cells. The proliferation and invasion abilities of tumor cells were detected to discuss the mechanism of Wnt/β-catenin signaling pathway in the enhanced malignant phenotype of non-small cell lung cancer induced by the anti-angiogenesis therapy. The specific siRNA could significantly inhibit the expression of VEGF in cells to simulate the anti-angiogenesis therapy. Under the action of 50nM VEGF siRNA, the proliferation ability of A549 significantly increased (P<0.05). After being treated with VEGF siRNA, the invasion ability of cells increased. Twenty-four hours after the transcription of 50nM siRNA into cells, the number of cells that come through the membrane was 278.3±12.9. Compared with the Ctrl siRNA group, when VEGF was inhibited, the expression of β-catenin and Cyclin D1 increased by 86% and 55% respectively. Meanwhile, the expression of E-cadherin decreased, while the one of vimentin increased. siRNA can significantly inhibit the expression of VEGF. For the anti-angiogenesis therapy, the inhibited expression of VEGF can activate the Wnt/β-catenin signaling pathway to cause the epithelial mesenchymal transition and then the enhanced malignant phenotype of non-small cell lung cancer.

Expression levels of vascular endothelial cell growth factor and microRNA-210 are increased in medulloblastoma and metastatic medulloblastoma.

The present study aimed to investigate the roles of the vascular endothelial cell growth factor (VEGF) and micro (mi)RNA-210 in the metastasis of primary medulloblastoma (MB) tumors. A total of 86 adult patients diagnosed with cerebellar MB were enrolled in the present study, of which 11 patients had metastatic MB in the subarachnoid space. The following samples were collected: MB primary tumor tissue, MB secondary tumor tissue, tumor adjacent tissues and cerebrospinal fluid (CSF). Immunohistochemical analyses of the tissue samples were conducted in order to detect patterns of VEGF expression. In addition, the expression levels of VEGF mRNA and miRNA-210 were analyzed using reverse transcription-quantititative polymerase chain reaction, and western blot analyses were used to investigate VEGF protein expression levels. The positive expression rate of VEGF was significantly higher in MB tumor tissue, as compared with adjacent tissues (P<0.01). In addition, VEGF mRNA and protein expression levels in MB primary and secondary tumor tissues, and in the CSF of patients with metastatic MB, were significantly upregulated, as compared with tumor adjacent tissues and the CSF of patients with non-metastatic MB, respectively (P<0.01). miRNA-210 expression levels were significantly upregulated in MB tumor tissues, the CSF of patients with metastatic MB and in tumor tissues of metastatic MB (P<0.01). In the present study, the expression levels of VEGF and miRNA-210 were upregulated in patients with MB and metastatic MB; thus suggesting that miRNA-210 may promote the metastasis of MB primary tumors by regulating the expression of VEGF.

Vascular endothelial growth factor-A and transforming growth factor-β1 expression in postmenopausal osteoporosis

Background Vascular endothelial growth factor (VEGF) plays an important role in bone angiogenesis, whereas transforming growth factor-β1 (TGF-β1) modulates bone differentiation, matrix formation, and cytokine expression. Aim of the work The aim of the study was to assess the immune expression of VEGF and TGF-β1 in cells lining the bone trabeculae among postmenopausal womεν in relation to bone histomorphometry and their association with osteoporosis. Patients and methods A total of 30 postmenopausal women were recruited for the study. Bone mineral density was measured using DEXA. On the basis of the results of the DEXA scan, they were divided as follows: group I comprised 14 (46.7%) women with normal bone mineral density; group II comprised five (16.7%) women with osteopenia; and group III comprised 11 (36.7%) women with osteoporosis. Iliac crest bone biopsy was taken from each of them. Histomorphometrical study was carried out using H&E stain to evaluate the trabecular microarchitecture in addition to immunohistochemistry for both VEGF and TGF-β1. Results Histomorphometric analysis was statistically and significantly different among the three groups. The area percentage of both TGF-β1 protein and VEGF expression in cells lining the bone trabeculae was statistically and significantly lower in the osteoporotic group compared with the other two groups. A statistically significant negative correlation was found between age and menopause duration and TGF-β1 only. A significant positive correlation was found between trabecular bone thickness and both VEGF (r=0.762, P=0.000) and TGF-β1 (r=0.785, P=0.000). Regression analysis showed only VEGF (t=2.537, P=0.018), and not TGF-β1 (t=0.326, P=0.747), to have independent association with trabecular bone thickness. Conclusion The results of the current study support that VEGF and TGF-β1 may contribute to the pathogenesis of postmenopausal osteoporosis. Further studies on the role of both TGF-β1 and VEGF in postmenopausal osteoporotic patients will be helpful to clarify the etiopathogenesis and treatment of postmenopausal osteoporosis.

Antimuscle atrophy effect of nicotine targets muscle satellite cells partly through an α7 nicotinic receptor in a murine hindlimb ischemia model

We have recently identified that donepezil, an anti-Alzheimer drug, accelerates angiogenesis in a murine hindlimb ischemia (HLI) model. However, the precise mechanisms are yet to be fully elucidated, particularly whether the effects are derived from endothelial cells alone or from other nonvascular cells. Further investigation of the HLI model revealed that nicotine accelerated angiogenesis by activation of vascular endothelial cell growth factor (VEGF) synthesis through nicotinic receptors in myogenic cells, that is, satellite cells, in vivo and upregulated the expression of angiogenic factors, for example, VEGF and fibroblast growth factor 2, in vitro. As a result, nicotine prevented skeletal muscle from ischemia-induced muscle atrophy and upregulated myosin heavy chain expression in vitro. The in vivo anti-atrophy effect of nicotine on muscle was also observed in galantamine, another anti-Alzheimer drug, playing as an allosteric potentiating ligand. Such effects of nicotine were attenuated in α7 nicotinic receptor knockout mice. In contrast, PNU282987, an α7 nicotinic receptor agonist, comparably salvaged skeletal muscle, which was affected by HLI. These results suggest that cholinergic signals also target myogenic cells and have inhibiting roles in muscle loss by ischemia-induced muscle atrophy.