Reduction Of Vascular Endothelial Growth Factor Research Articles

Expression Level of IL20, mTOR, IL13RA1, and SEMA4C Are Increased in the Visceral Adipose Tissue of Patients With Non-Alcoholic Steathepatitis While There is a Decrease in the Expression Levels of Their Regulatory Mirnas

fully understood. We hypothesized that probiotics could strengthen barrier function in an animal model of alcoholic liver disease (ALD) by activating hypoxia-inducible factor (HIF) leading to an increase in barrier-protecting factors, such as intestinal trefoil factor (ITF), vascular endothelial growth factor (VEGF), and CD73, and thus prevent endotoxemia and subsequently ALD. Male C57/6J mice were fed Lieber DiCarli diet containing 5% ethanol for 8 weeks. In last two weeks, 1 x 109 cfu/daily live Lactobacillus GG (LGG) was added to the diet. Serum endotoxin and alanine aminotransferase were measured. Intestinal and liver tissues were analyzed for ITF, VEGF and CD73. In addition, liver steatosis was assessed by histology. The mice treated with LGG had significantly less hepatic steatosis, serum ALT and lipopolysaccharide levels, and less intestinal barrier damage than alcohol-fed mice. ITF, VEGF and CD73 were deceased in the intestine of mice treated with alcohol, and LGG attenuated this decrease. In Vitro studies revealed that treatment with 5% alcohol decreased transepithelial resistance and increased in FITC-dextran permeability in Caco-2 cells. Supplementation of LGG cultural supernatant (LGGs) attenuated these deleterious effects in a timedependent manner. LGGs also prevented alcohol-induced reduction in ITF, VEGF, and CD73 which are known HIF-1 targets. These results indicate that LGG attenuating the alcohol-induced liver injury is likely, at least in part, by activating HIF signaling leading to intestinal mucosal barrier protection.

Inhibition of tumor growth by quercetin with increase of survival and prevention of cachexia in Walker 256 tumor-bearing rats

Quercetin, a flavonoid abundantly present in fruit, vegetables, wine and tea, has revealed several properties such as antioxidant, antiproliferative and anticancer. Cachexia is a poorly understood syndrome present in already compromised cancer patients, decreasing the quality of life and increasing mortality. Many studies have been performed in an attempt to discover an effective treatment for cachexia, but none of the tested therapies has fulfilled expectations. The objective of the present study was to analyze the effect of quercetin in the therapeutic treatment of cachexia and reversion of tumor growth in rats bearing Walker 256 carcinosarcoma (W256). Rats bearing W256 were treated daily with I.P. quercetin injections, at different doses (10, 15, 25 and 35 mg/kg). The results show that 10 mg/kg quercetin inhibited tumor growth by about 50% (ED 50) when compared with controls (CTR). Moreover, two animals of this group presented complete tumor regression. Matrix metalloproteinase-2 (MMP-2) activity and vascular endothelial growth factor (VEGF) expression decreased in rats bearing W256 treated with 10 mg/kg quercetin when compared with CTR. Thus, the inhibition of tumor growth, survival increase, decrease of MMP-2 and VEGF levels and reduction of cachexia in animals treated with quercetin strongly support the anticancer function of this flavonoid.

Effects of Spironolactone and Losartan on Diabetic Nephropathy in a Type 2 Diabetic Rat Model

BackgroundWhile there is an evidence that the anti-inflammatory properties of spironolactone can attenuate proteinuria in type 2 diabetes, its effects on vascular endothelial growth factor (VEGF) expression in diabetic nephropathy have not been clearly defined. In this study, we examined the effects of spironolactone, losartan, and a combination of these two drugs on albuminuria, renal VEGF expression, and inflammatory and oxidative stress markers in a type 2 diabetic rat model.MethodsThirty-three Otsuka-Long-Evans-Tokushima-Fatty (OLETF) rats were divided into four groups and treated with different medication regimens from weeks 25 to 50; OLETF diabetic controls (n=5), spironolactone-treated (n=10), losartan-treated (n=9), and combination of spironolactone- and losartan-treated (n=9).ResultsAt week 50, the albumin-to-creatinine ratio was significantly decreased in the losartan and combination groups compared to the control OLETF group. No decrease was detected in the spironolactone group. There was a significant reduction in renal VEGF, transforming growth factor (TGF)-β, and type IV collagen mRNA levels in the spironolactone- and combination regimen-treated groups. Twenty-four hour urine monocyte chemotactic protein-1 levels were comparable in all four groups but did show a decreasing trend in the losartan and combination regimen groups. Twenty-four hour urine malondialdehyde levels were significantly decreased in the spironolactone- and combination regimen-treated groups.ConclusionThese results suggest that losartan alone and a combined regimen of spironolactone and losartan could ameliorate albuninuria by reducing renal VEGF expression. Also, simultaneous treatment with spironolactone and losartan may have protective effects against diabetic nephropathy by decreasing TGF-β and type IV collagen expression and by reducing oxidative stress in a type 2 diabetic rat model.

HO-3867, a Synthetic Compound, Inhibits the Migration and Invasion of Ovarian Carcinoma Cells through Downregulation of Fatty Acid Synthase and Focal Adhesion Kinase

Fatty acid synthase (FAS) and focal adhesion kinase (FAK), which are overexpressed in a variety of human epithelial tumors, play a key role in the migration and invasion of cancer cells. Hence, strategies targeted at inhibiting the FAS/FAK proteins may have therapeutic potential for cancer treatment. The goal of the present study was to determine the effect of HO-3867, a synthetic compound, on the migratory ability of ovarian cancer cells and to understand the mechanistic pathways including the involvement of FAS, FAK, and associated signaling proteins. The study was done using two established human ovarian cancer cell lines, A2780 and SKOV3. Incubation with 10 μmol/L HO-3867 for 24 hours significantly inhibited the native as well as the vascular endothelial growth factor (VEGF)-mediated migration and invasion of the cells. HO-3867 significantly attenuated FAS and FAK protein levels apparently through accelerated ubiquitin-dependent degradation, as shown by a clear downregulation of isopeptidase USP2a. Exposure of cells to HO-3867 also significantly inhibited FAS activity and mRNA levels and a number of downstream proteins, including phospho-extracellular signal-regulated kinase 1/2, phospho-human epidermal growth factor receptor 1, sterol regulatory element binding protein 1, VEGF, and matrix metalloproteinase 2. Western blot and immunohistochemical analyses of A2780 xenograft tumors in mice treated with HO-3867 showed significant reduction in FAS, FAK, VEGF, and downstream protein levels when compared with the untreated control. Collectively, the results showed that HO-3867 suppressed the migration and invasion of ovarian cancer cells by inhibiting the expression or activity of FAS and FAK proteins. The study suggests that molecular targeting of FAS and FAK by HO-3867 may be a potential strategy for ovarian cancer therapy.

Evaluation of the Role of Vascular Endothelial Growth Factor in Diabetic Retinopathy

Purpose: The study is aiming at investigating the correlation between vascular endothelial growth factor (VEGF) as an angiogenic factor and diabetic retinopathy in type 2 diabetic patients and the effect of panretinal coagulation and glycemic control on VEGF. Methods: The study included 30 patients of type 2 diabetes, 10 of them did not suffer from any of the vascular complications of type 2 diabetes mellitus (group 1), 10 patients had nonproliferative diabetic retinopathy (group 2), 10 patients had proliferative diabetic retinopathy (PDR) (group 3) and (group 4), as well as 10 healthy subjects that served as control group. All participants were subjected to complete clinical examination including ophthalmic and medical examination, laboratory investigations comprising complete blood count, liver function test, serum creatinine, 24-hour urinary albumin excretion, lipid profile, fasting and 2-hour postprandial blood glucose, HbA_1C and serum VEGF. Results: The study reported a highly significant increase in the serum VEGF in the diabetic patients compared to the control group (p < 0.001), and there was also a highly significant increase in the serum VEGF in the patients with PDR versus nonproliferative diabetic retinopathy (40.55 ± 8.28 vs. 20.3 ± 2.45, p < 0.001). There was a reduction in the serum VEGF in a group of diabetic patients with poor glycemic control when their diabetic state corrected through 4 months of follow-up was highly significant (17.29 ± 1.61 before vs. 9.39 ± 0.82 after control p < 0.001) as well as the reduction in the serum VEGF which was observed in a group of patients with PDR when proper panretinal photocoagulation (PRP) was applied to their retinae with 4 months of follow-up (40.55 ± 8.28 before vs. 21.15 ± 1.76 after PRP, p < 0.001). Conclusion: Serum VEGF is significantly increased in diabetic patients, especially those with PDR, and this elevation of VEGF was reduced in uncontrolled diabetic patients with proper gycemic control, and in patients with PDR with proper PRP, indicating that VEGF is an angiogenic factor that reflects the degree of neovascularization in diabetic complications.

What Type of VEGF Do You Need?

Typically considered a mediator of angiogenesis and vascular permeability, vascular endothelial growth factor (VEGF) has emerged as an important modulator of glomerular health and disease. The pleiotropic nature VEGF was dramatically illustrated in cases in which patients who were undergoing chemotherapy with VEGF antibodies ( e.g. , bevacizumab) developed proteinuria1 and thrombotic microangiopathy,2 indicating damage to the glomerular filtration barrier. In addition, individuals with preeclampsia, a condition characterized by excess circulating levels of a soluble form of the VEGF receptor, can likewise develop proteinuria,3 and increased serum levels of sFlt may contribute to endothelial dysfunction in chronic kidney disease.4 These studies suggest that a threshold of VEGF must be met to ensure glomerular health. Accordingly, in this editorial space just 4 years ago, Mathieson asked, “How much VEGF do you need?”5 At the time, the question was prompted by several studies demonstrating that VEGF gene dose is critical to glomerular development, structure, and function. Quaggin's group6,7 was reporting the generation of a series of gene-targeted mice with podocyte-specific VEGF reduction, with expression ranging from 0 to 50% of normal. This exquisite control over VEGF expression yielded a striking variety of phenotypes: When VEGF expression is ablated entirely, glomerular development fails and perinatal lethality ensues; low expression levels (approximately 25% of normal) results in a cascade of glomerular cell attrition beginning with endothelial and proceeding to mesangial cells shortly after birth, and intermediate VEGF levels (approximately 50% of normal) recapitulates …

The role of vascular endothelial growth factor in the progression of diabetic vascular complications

This study was planned to study the relationship between vascular endothelial growth factor (VEGF) as an angiogenic factor and different micro- and macrovascular complications in type II diabetic patients and look for a possible role of control on the serum level of VEGF. The study included 55 type II diabetic patients, 10 of them were not complicated with any of the vascular complications of type II diabetes mellitus (DM) (group 1), 21 patients had microvascular complication either retinopathy, nephropathy, or neuropathy (group 2), 14 patients had macrovascular complications either coronary artery disease or peripheral vascular disease (group 3), and 10 patients with mixed micro- and microvascular complications (group 4), as well as 15 healthy subjects served as control group. All subjects were subjected to complete clinical examination, including fundus examination, proper investigations with stress on electrocardiography, electromyography, nerve conduction velocity, Doppler study of the peripheral arteries, and laboratory investigations such as complete blood count, liver function test, serum creatinine, 24-h urinary albumin excretion, lipid profile, fasting and 2-h postprandial blood glucose, glycosylated haemoglobin (HbA(1c)), and serum VEGF. The study revealed that there was a highly significant increase in the serum VEGF in the diabetic patients compared with the control group. The P-value (P<0.001) was detected and there was also a highly significant increase in the serum VEGF in the patients with different micro- and macrovascular diabetic complications compared with uncomplicated diabetic group (P<0.001). A highly significant increase in the serum VEGF in diabetic patients with proliferative diabetic retinopathy was detected compared with non-proliferative diabetic retinopathy (40.55±8.28 vs20.3±2.45, P<0.001) and in diabetic nephropathic patients with macroalbuminuria compared with those with microalbuminuria (36.14±6.99 vs19.42±2.44, P<0.001). The reduction of the serum VEGF in a group of diabetic patients with poor control when their diabetic state was corrected through 4 months follow-up was highly significant (17.29±1.61 before vs9.39±0.82 after control P<0.001), as well as the reduction of the serum VEGF, which was observed in a group of patients with proliferative diabetic retinopathy (PDR) when proper pan retinal photocoagulation (PRP) was applied to their retinae with 4 months follow-up (40.55±8.28 before vs21.15±1.76 after PRP, P<0.001). On the other hand, the impact of some clinical and laboratory parameters of our diabetic patients on the serum VEGF revealed significant positive correlations between serum VEGF and age of the patients, duration of diabetes, systolic and diastolic blood pressure, body mass index, fasting and 2-h postprandial blood glucose, HbA(1c), serum creatinine, degree of albuminuria and total cholesterol, LDL, and platelet count. Serum VEGF was significantly increased in diabetic patients especially with micro- and macrovascular complications and the proper control of diabetes reduced the elevation of serum VEGF in uncontrolled diabetic patients, and in patients with PDR with proper PRP, indicating that VEGF is an angiogenic factor that reflects the degree of neovascularization in diabetic complications.

Effects of Chronic Red Wine Consumption on the Expression of Vascular Endothelial Growth Factor, Angiopoietin 1, Angiopoietin 2, and Its Receptors in Rat Erectile Tissue

Long-term consumption of red wine (RW) apparently confers some protection against cardiovascular diseases due to antiatherosclerotic properties of polyphenols and ethanol (EtOH). There is some evidence indicating that they do so by regulating angiogenesis, but the mechanism and the modulator factors involved are largely unknown. The aim of this study was to evaluate the effects of chronic ingestion of RW in vascular structure and in the pattern of expression of vascular growth factors in the rat corpus cavernosum. Male Wistar rats aged 6 mo were treated with RW or an equivalent EtOH solution, as the only liquid source for 6 mo. Expression of vascular endothelial growth factor (VEGF), angiopoietin 1 and angiopoietin 2, and their receptors (VEGFR1, VEGFR2, and Tie2) in cavernous tissue was assayed by immunofluorescence and Western blotting. A reduction of VEGF and VEGFR2 expression, respectively, in smooth muscle and endothelial cells was observed in RW-treated animals, which was balanced by an increase in angiopoietins/Tie2 expression. In EtOH rats, only a decrease in expression of the receptors VEGFR2 and Tie2 was observed. These results, taken together, suggest that antioxidants present in RW activate selected mechanisms for the maintenance of cavernous tissue vascularization. However, functional studies will be necessary to elucidate if RW is of benefit in the prevention of deleterious vascular events associated with ED.

Effect of pan-retinal laser photocoagulation on plasma VEGF, endothelin-1 and nitric oxide in PDR.

To study plasma levels of vascular endothelial growth factor (VEGF), endothelin-1(ET-1) and nitric oxide (NO) in patients with proliferative diabetic retinopathy (PDR) before and after pan-retinal photocoagulation (PRP). Forty patients (23 females and 17 males, mean age 48.5±12.2 years) with PDR without previous PRP therapy were studied. Blood samples were obtained before and 3 months after the last PRP session. Baseline (prelaser) plasma levels of VEGF, ET-1 and NO were compared with their levels in 30 healthy age- and sex- matched controls and also with plasma levels 3 months post-PRP. Patients with PDR had significantly raised plasma VEGF (375±89ng/L), ET-1(20±5ng/L) and NO (135±53µmol/L) when compared with healthy control group (P<0.01). After PRP, there was a significant reduction in plasma VEGF (179±66ng/L), ET-1 (11±5ng/L) and NO (91±49µmol/L) levels at 3 months' follow-up but still significantly higher than healthy controls. Patients with PDR demonstrate elevated VEGF, ET-1 and NO, which decrease after successful laser treatment.

Metabolic programming of ovarian angiogenesis and folliculogenesis by maternal malnutrition during lactation

To evaluate whether maternal malnutrition during lactation programs ovarian folliculogenesis and the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) and its receptors KDR, Flt-1, and FGFR. Experimental study. University-based research laboratory. Adult female rats from a urogenital research laboratory. Six rat dams randomly assigned to the following groups: control group (C), with free access to a standard laboratory diet containing 23% protein; and a protein-energy-restricted group (PER), with free access to an isoenergy and protein-restricted diet containing 8% protein. After weaning, the female pups had free access to the standard laboratory diet until 90 days of age, when they were sacrificed at the proestrum stage. Quantification of ovarian follicles, vessels, and expression of growth factors and their receptors. Maternal malnutrition during lactation caused a significant reduction in the number of primordial (C = 6.60 +/- 0.24, PER = 5.20 +/- 0.20), primary (C = 5.80 +/- 0.66, PER = 4.00 +/- 0.31), and Graafian follicles/section (C = 2.18 +/- 0.29, PER = 1.08 +/- 0.37), in KDR (C = 0.22 +/- 0.04, PER = 0.09 +/- 0.01), Flt-1 (C = 0.28 +/- 0.05, PER = 0.12 +/- 0.02), and FGFR mRNA expression (C = 0.34 +/- 0.05, PER = 0.13 +/- 0.05) and in the vessel density of follicles (C = 17.26 +/- 2.30, PER = 9.96 +/- 0.97). Maternal malnutrition during lactation programs the follicular development by a reduction of VEGF and FGF mRNA receptors expression, probably from a direct action on the follicular development or a reduction in vasculature resulting in a decreased delivery of folliculotrophic substances in PER animals.

Chondrogenic Priming of Human Bone Marrow Stromal Cells: A Better Route to Bone Repair?

The use of bioengineered cell constructs for the treatment of bone defects has received much attention of late. Often, bone marrow stromal cells (BMSCs) are used that are in vitro-stimulated toward the osteogenic lineage, aiming at intramembranous bone formation. The success of this approach has been disappointing. A major concern with these constructs is core degradation and necrosis caused by lack of vascularization. We hypothesized that stimulation of cells toward the endochondral ossification process would be more successful. In this study, we tested how in vitro priming of human BMSCs (hBMSCs) along osteogenic and chondrogenic lineages influences survival and osteogenesis in vivo. Scaffolds that were pre-cultured on chondrogenic culture medium showed collagen type II and collagen type X production. Moreover, vessel ingrowth was observed. Priming along the osteogenic lineage led to a mineralized matrix of poor quality, with few surviving cells and no vascularization. We further characterized this process in vitro using pellet cultures. In vitro, pellets cultured in chondrogenic medium showed progressive production of collagen type II and collagen type X. In the culture medium of these chondrogenic cultured pellets, vascular endothelial growth factor (VEGF) release was observed at days 14, 21, and 35. When pellets were switched to culture medium containing beta-glycerophosphate, independent of the presence or absence of transforming growth factor beta (TGF-beta), mineralization was observed with a concomitant reduction in VEGF and matrix metalloproteinase (MMP) release. By showing that VEGF and MMPs are produced in chondrogenically differentiated hBMSCs in vitro, we demonstrated that these cells produce factors that are known to be important for the induction of vascularization of the matrix. Inducing mineralization in this endochondral process does, however, severely diminish these capacities. Taken together, these data suggest that optimizing chondrogenic priming of hBMSCs may further improve vessel invasion in bioengineered constructs, thus leading to an alternative and superior approach to bone repair.

Angiotensin-(1–7), a small molecule inhibitor of glioblastoma growth

2024 Background: Recent studies suggest that anti-angiogenic therapies may be effective in patients with glioblastoma multiforme (GBM), a highly vascular tumor. We have previously demonstrated that angiotensin-(1–7) [Ang-(1–7)], an endogenous heptapeptide with anti-angiogenic properties, significantly reduced the serum-stimulated growth of three human glioblastoma cell lines which contained mRNA for the AT(1–7) receptor mas. We now provide the first evidence that Ang-(1–7) markedly decreases the proliferation and growth of human glioblastomas in vivo using a xenograft model. Methods: Athymic mice with tumors resulting from injection of human U87 glioblastoma cells were treated for 18 days with saline or 1000 μg/kg Ang-(1–7), delivered by subcutaneous injection every 12 h. Results: The average volume of the tumors from mice treated with the heptapeptide was approximately 3-fold less than the size of the tumors from control animals (586.1 ± 94.5 mm3 vs 1845.5 ± 238.1 mm3; n = 6, p &lt; 0.05). Further, the tumors from mice injected with Ang-(1–7) weighed almost 50% less than the tumors from mice treated with saline (1.31 ± 0.12 g vs. 2.45 ± 0.23 g; n = 6, p &lt; 0.05). The Ang-(1–7)-mediated reduction in tumor growth was associated with a significant decrease in immunoreactive Ki67, a proliferation marker. In addition, a marked down-regulation of cyclooxygenase 2 (COX-2), prostaglandin E synthase (PGES-1) and vascular endothelial growth factor (VEGF) was observed in tumors from Ang-(1–7)-injected mice compared to saline-treated controls (COX-2: 1.00 ± 0.06 vs 0.55 ± 0.07 relative gene expression; PGES-1: 1.03 ± 0.08 vs. 0.40 ± 0.06; VEGF: 1.05 ± 0.07 vs. 0.59 ± 0.09; n = 6, p &lt; 0.05) with no effect on COX-1 or PGI synthase mRNA. Conclusions: These results suggest that Ang-(1–7) may reduce the concentration and ratio of proliferative and anti-proliferative prostaglandins to decrease glioblastoma growth as well as attenuate angiogenesis through a reduction in VEGF. Thus, Ang-(1–7) may be a new, first-in-class small molecule inhibitor for the treatment of glioblastoma, providing combination therapy as a selective COX-2/PGES-1 and angiogenic inhibitor, targeting a specific AT(1–7) receptor mas. No significant financial relationships to disclose.

Anti-angiogenic activity of carebastine: a plausible mechanism affecting airway remodelling

Ebastine is a well-known selective second-generation histamine H(1) receptor antagonist, which is used for seasonal and perennial allergic rhinitis and chronic urticaria. Angiogenesis plays a crucial role in the development of airway inflammation and remodelling in allergic rhinitis and asthmatic patients, in whom, indeed, the mucosa displays increased vascularity and overexpression of vascular endothelial growth factor (VEGF). The aim of the present study was to evaluate the anti-angiogenic properties of carebastine, the active metabolite of ebastine. The effects of carebastine on human umbilical vein endothelial cell (EC) (HUVEC) and human pulmonary artery EC (HPAEC) proliferation, migration and capillary-like tube formation were investigated in vitro, and in the chick embryo chorioallantoic membrane (CAM) assay in vivo. Moreover, the effect of carebastine on phosphorylation of the cell VEGF receptor fetal liver kinase-1, or VEGF receptor 2 (VEGFR-2), and Akt kinase (Akt) was evaluated by Western blotting. Carebastine inhibited VEGF-induced HUVEC and HPAEC proliferation, migration and angiogenesis in a dose-dependent manner in vitro. Cell proliferation was inhibited by 42 and 64% in HUVECs and 62 and 75% in HPAECs upon exposure for 48 and 72 h, respectively, to 20 microM carebastine (p < or = 0.03), and even more with 30 microM carebastine. Cell migration was inhibited by 37 and 70% in HUVECs (p < or = 0.03) and 60 and 78% in HPAECs (p < or = 0.01) in the presence of 10 and 30 microM carebastine, respectively. Carebastine (20 microM) caused a significant reduction (70-86%; p<0.01) in topological parameters of the capillary network produced in vitro by both EC lines on a basement membrane extract. Carebastine (30 and 50 microM) inhibited the VEGF-induced angiogenesis in the CAM assay in vivo two- and three-fold, respectively (p<0.001). Finally, both EC lines, on exposure to 10 and 20 microM carebastine, showed a four- to six-fold reduction (p < or = 0.01) in both VEGF- and H1 receptor-induced VEGFR-2 and Akt phosphorylation. Overall, these data provide the first evidence regarding the anti-angiogenic activity of ebastine, and suggest its potential use as an anti-angiogenic molecule, besides its antihistaminic activity for the treatment of allergic diseases in which angiogenesis takes place.

Reduction in VEGF Protein and Phosphorylated Nephrin Associated with Proteinuria in Adriamycin Nephropathy Rats

Background/Aims: The relationship between vascular endothelial growth factor (VEGF) and the phosphorylated critical podocyte slit diaphragm molecule nephrin is not fully clarified. This study investigated the dynamic changes in VEGF expression and nephrin phosphorylation, and the effects of the antiproteinuric drugs prednisone and lisinopril on them in Adriamycin nephropathy rats. Methods: Renal tissues from Adriamycin rats were collected at days 3, 7, 14, and 28. Distribution and expression of VEGF was revealed by immunohistochemistry, real-time PCR and Western blot. Phosphorylated nephrin was evaluated by immunoprecipitation. Results: A discontinuous redistribution of VEGF was displayed at day 3, followed by significant protein reduction at day 7 with persistent downregulation to day 28. Phosphorylated nephrin decreased evidently at day 14 and persisted to day 28. The reduction in VEGF and phosphorylated nephrin was not a result of podocyte loss. The intervention of prednisone and lisinopril evidently reduced proteinuria, effectively attenuated the severe lesions of podocyte foot processes, and restored the reduction in VEGF and nephrin phosphorylation. At day 28, the reduction in VEGF and phosphorylated nephrin was negatively correlated with proteinuria, whereas the phosphorylated nephrin was positively correlated with VEGF protein from day 7 to day 28. Conclusion: The reduction in VEGF protein and nephrin phosphorylation was possibly involved in the proteinuria in Adriamycin rats, and there might be some relationship between VEGF and nephrin phosphorylation. The antiproteinuric effects of lisinopril and prednisone were achieved at least partially by restoring VEGF protein and nephrin phosphorylation.

HIV-1-mediated delivery of a short hairpin RNA targeting vascular endothelial growth factor in human retinal pigment epithelium cells

Background:Vascular endothelial growth factor (VEGF) has been shown to play a major role in the pathological neovascularisation that occurs in degenerative retinal diseases like age-related macular degeneration (AMD). Although several...

GROWTH FACTORS DECREASE IN SUBJECTS WITH MILD TO MODERATE ALZHEIMER'S DISEASE (AD): POTENTIAL CORRECTION WITH DEHYDROEPIANDROSTERONE-SULPHATE (DHEAS)

The integrity of neuroprotection is an important component against the development of cognitive disorders and AD. Within this context, DHEAS would seem to have some positive metabolic and endocrine effects to delay brain aging by recovering the impairment of neuroprotective growth factors. In the present study we measured by ELISA the secretion of insulin-like growth factor-1 (IGF-1), vascular endothelial growth factor (VEGF), and transforming growth factor-β1 (TGFβ1) in the supernatants of cultured circulating peripheral blood mononuclear cells (PBMC) from which natural killer cells (NK) were separated (PBMC-NK) (pg/ml/7.75 × 10 6 cells) in healthy subjects and in age-matched patients with mild to moderate AD. The growth factors were measured in spontaneous conditions and after stimulation with growth hormone (GH) 1 μg/ml (IGF-1), lipopolysaccharide (LPS) 1 μg/ml (VEGF) and glucose 10 μM (TGF(β1). AD group demonstrated at baseline a severe reduction of IGF-1 (3.7 + 1.2 pg/ml after GH), VEGF (63 ± 18 pg/ml spontaneous and 210 ± 65 pg/ml after LPS) and TGF(β1 (33 ± 10 pg/ml spontaneous and 75 ± 12 pg/ml after glucose) secretions compared to healthy elderly subjects (IGF-1, 9.5 ± 2.8 pg/ml after GH, p < 0.001; VEGF, 117 ± 38 pg/ml spontaneous, p < 0.001 and 690 ± 120 pg/ml after LPS, p < 0.001; and TGF(β1, 73 ± 21 pg/ml spontaneous, p < 0.001 and 169 ± 53 pg/ml after glucose, p < 0.001). Significant positive correlations between IGF-1 and VEGF concentrations were found both in healthy subjects (r = 0.87, p < 0.001) and in AD subjects (r = 0.87, p < 0.001). The co-incubation of NK cells with DHEAS (10 6 M/ml/cells) significantly increase IGF-1, VEGF and TGF (β1 production, reaching in AD group the normal concentrations found in healthy subjects (IGF-1, 7.9 + 2.4 pg/ml after GH; VEGF, 105 ± 31 pg/ml spontaneous and 670 ± 112 pg/ml after LPS; and TGFfβ1, 68 ± 18 pg/ml spontaneous and 155 ± 48 pg/ml after glucose). These data suggested that DHEAS is able to increase the immunoendocrine production of neuroprotective growth factors, which is reduced in AD subjects, so suggesting a new approach in the treatment of dementia.

A Somatostatin Analogue, Octreotide, Inhibits the Occurrence of Second Primary Tumors and Lung Metastasis after Resection of Hepatocellular Carcinoma in Mice

Occurrence of second primary tumors and metastasis remains the major obstacles to prolonged survival of patients with primary hepatocellular carcinoma (HCC). A somatostatin analogue, octreotide, has been previously reported to inhibit the growth of human HCC xenografts in nude mice through its anti-angiogenic activity. It is therefore important to investigate whether octreotide could prevent second primary hepatomas or distant metastasis following resection of primary HCC. In this study, nude mice, bearing the human HCC xenografts with highly metastatic potential (LCI-D20) in the left liver lobe, underwent tumor resection, and received intraperitoneal administration of octreotide or saline as a control for 35 consecutive days. Compared with the control group, octreotide at the doses of 100 and 200 microg/kg/day significantly inhibited the growth (P < 0.001 and P < 0.001, respectively) and incidence of second primary tumors (P = 0.016 and P = 0.001, respectively), decreased lung metastasis (P < 0.001 and P < 0.001, respectively), and prolonged the life span (P = 0.029 and P = 0.006, respectively). Moreover, intratumoral angiogenesis quantified by microvessel density as well as serum and tissue vascular endothelial growth factor (VEGF) levels were considerably decreased in octreotide-treated animals compared to the control animals. These findings suggest that octreotide may prevent the occurrence of second primary hepatomas and lung metastasis after resection of primary HCC, which may be partially attributed to down-regulation of VEGF and subsequent reduction in tumor angiogenesis. Octreotide administration may be useful as an adjuvant therapy to improve survival of patients with HCC.

Chondrogenic Priming of Human Bone Marrow Stromal Cells: A Better Route to Bone Repair?

The use of bioengineered cell constructs for the treatment of bone defects has received much attention of late. Often, bone marrow stromal cells (BMSCs) are used that are in vitro-stimulated toward the osteogenic lineage, aiming at intramembranous bone formation. The success of this approach has been disappointing. A major concern with these constructs is core degradation and necrosis caused by lack of vascularization. We hypothesized that stimulation of cells toward the endochondral ossification process would be more successful. In this study, we tested how in vitro priming of human BMSCs (hBMSCs) along osteogenic and chondrogenic lineages influences survival and osteogenesis in vivo. Scaffolds that were pre-cultured on chondrogenic culture medium showed collagen type II and collagen type X production. Moreover, vessel ingrowth was observed. Priming along the osteogenic lineage led to a mineralized matrix of poor quality, with few surviving cells and no vascularization. We further characterized this process in vitro using pellet cultures. In vitro, pellets cultured in chondrogenic medium showed progressive production of collagen type II and collagen type X. In the culture medium of these chondrogenic cultured pellets, vascular endothelial growth factor (VEGF) release was observed at days 14, 21, and 35. When pellets were switched to culture medium containing beta-glycerophosphate, independent of the presence or absence of transforming growth factor beta (TGF-beta), mineralization was observed with a concomitant reduction in VEGF and matrix metalloproteinase (MMP) release. By showing that VEGF and MMPs are produced in chondrogenically differentiated hBMSCs in vitro, we demonstrated that these cells produce factors that are known to be important for the induction of vascularization of the matrix. Inducing mineralization in this endochondral process does, however, severely diminish these capacities. Taken together, these data suggest that optimizing chondrogenic priming of hBMSCs may further improve vessel invasion in bioengineered constructs, thus leading to an alternative and superior approach to bone repair.

Effect of arsenic trioxide on proliferation and apoptosis of U266 cells and its relationship with the expression variation of VEGF

The aim of this article is to explore the effect of arsenic trioxide (As2O3) on the proliferation and apoptosis of myeloma cell line U266 and its relationship with the expression variation of vascular endothelial growth factor (VEGF). The viability and apoptosis of U266 cells were observed by methylthiazolyl-tetrazolium (MTT) assay and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The effect of As2O3 on the VEGF expression of U266 cells were tested by enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) analysis. We found that As2O3 could significantly inhibit the growth of U266 cells, and the concentration for 50% growth inhibition (IC50) was 2 μmol/L. After treatment with 2, 5, 10 μmol/L As2O3 for 36 hours, dose-dependent apoptosis of U266 cells was observed. After treatment with 2, 5, 10 μmol/L As2O3 for 72 hours, a dose-dependent reduction of VEGF in the supernatant of U266 cells culture was found. As far as single cells are concerned, nevertheless, the expression of VEGF mRNA did not vary. So we draw the conclusion that As2O3 could induce the apoptosis of U266 cells and inhibit their proliferation, decrease the tumor load, and lead to the reduction of VEGF in the culture supernatant, but not change the expression of VEGF in single U266 cells.

Vascular Endothelial Growth Factor production in breast cancer is reduced by Camptothecin regardless of Cyclooxygenase-2 expression

Introduction: Breast cancer metastasis requires angiogenesis. Cyclooxygenase-2 (COX-2) and Vascular Endothelial Growth Factor (VEGF) are implicated in this process. Previous reports suggest blocking the COX-2 pathway inhibits angiogenesis via reduction of VEGF. This study investigated the effect of two compounds on COX-2 and VEGF in breast cancer.