Suppression of Human Tenon Fibroblast Cell Proliferation by Lentivirus-Mediated VEGF Small Hairpin RNA

Purpose. The functions of vascular endothelial growth factor (VEGF) in scar formation after trabeculectomy were investigated in a human Tenon fibroblast cell line from glaucoma patients using lentivirus-mediated VEGF shRNA. Methods. Human Tenon fibroblast (HTF) cells were isolated from scar tissue of glaucoma patients during secondary surgery. Lentivirus-VEGF-shRNA was constructed and transfected into HTF cells. Subsequently, VEGF mRNA and protein expression were analyzed using quantitative RT-PCR and western blotting, respectively, and the effects of VEGF knockdown were analyzed. The inhibition of HTF proliferation was monitored according to total cell numbers using ScanArray. Results. Both mRNA and protein levels of VEGF were reduced by lentivirus-mediated VEGF-shRNA, and proliferation of HTF cells was inhibited. Conclusions. Primary cultures of human Tenon fibroblast (HTF) were established, and proliferation was decreased following inhibition of VEGF. VEGF may be a suitable therapeutic target for reducing scar tissue formation in glaucoma patients after filtration surgery.

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In Vitro Study on the Regulation of Annexin IV and VEGF by hCG in the Human Endometrium

Biochemistry Research International ◽

10.1155/2020/8892930 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Shaoyuan Xu ◽

Jie Li ◽

Xiaoyan Chen ◽

Beiyu Liu

Keyword(s):

Epithelial Cells ◽

In Vitro Study ◽

Control Group ◽

Vascular Endothelial ◽

Vegf Mrna ◽

Endometrial Cells ◽

Mrna And Protein Expression ◽

Endothelial Growth Factor ◽

Annexin Iv

Objective. Whether changes in vascular endothelial growth factor (VEGF) and annexin IV during implantation are regulated through the LH/hCG-R needs further research. To investigate the mechanism of hCG on the expression of annexin IV and VEGF in human endometrial cells. Methods. Endometrial cells were isolated and identified from human specimens. The proportion of glandular and epithelial cells was analyzed. Annexin IV and VEGF were analyzed by qRT-PCR (mRNA), western blot (proteins), and immunohistochemistry (proteins). Protein location was identified by immunohistochemistry. The cells were cultured with hCG, hCG/PD98059 (a MAPK inhibitor), or no treatment (control). Results. The proportions between the glandular epithelial cells and stromal cells at inoculation and when adding hCG were 25.8 ± 0.2% and 27.8 ± 0.04%, respectively ( P > 0.05 ). LH/hCG-R, annexin IV, and VEGF were found in the cytoplasm of endometrial cells. After 2, 6, 12, and 24 h of hCG treatment, compared with 1 h, VEGF mRNA was increased by 1.25-fold, 3.19-fold, 4.21-fold, and 4.86-fold and annexin IV by 2.23-fold, 3.37-fold, 5.14-fold, and 5.02-fold. Compared with the control group, annexin IV mRNA and protein were increased in the hCG and hCG/PD98059 groups (mRNA/protein: 1.99-fold/1.80-fold and 2.33-fold/1.93-fold, P < 0.05 ). Compared with the control group, VEGF mRNA and protein were increased in the hCG group (mRNA/protein: 2.30-fold/1.86-fold), but not in the hCG/PD98059 group. Conclusion. hCG could upregulate the mRNA and protein expression of annexin IV and VEGF. The upregulation of annexin IV by hCG could not be inhibited by PD98059, but the upregulation of VEGF by hCG could.

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Proangiogenic properties of human myeloma cells: production of angiopoietin-1 and its potential relationship to myeloma-induced angiogenesis

Blood ◽

10.1182/blood-2002-10-3257 ◽

2003 ◽

Vol 102 (2) ◽

pp. 638-645 ◽

Cited By ~ 73

Author(s):

Nicola Giuliani ◽

Simona Colla ◽

Mirca Lazzaretti ◽

Roberto Sala ◽

Giovanni Roti ◽

...

Keyword(s):

Myeloma Cell ◽

Vascular Endothelial ◽

Myeloma Cells ◽

Vessel Formation ◽

Protein Levels ◽

Mrna And Protein Expression ◽

Endothelial Growth Factor ◽

Angiopoietin 1

AbstractPatients with multiple myeloma (MM) have increased bone marrow (BM) angiogenesis; however, the proangiogenic properties of myeloma cells and the mechanisms of MM-induced angiogenesis are not completely clarified. The angiopoietin system has been identified as critical in the regulation of vessel formation. In this study we have demonstrated that myeloma cells express several proangiogenic factors, and, in particular, we found that angiopoietin-1 (Ang-1), but not its antagonist Ang-2, was expressed by several human myeloma cell lines (HMCLs) at the mRNA and the protein levels. In a transwell coculture system, we observed that myeloma cells up-regulated the Ang-1 receptor Tie2 in human BM endothelial cells. Moreover, in an experimental model of angiogenesis, the conditioned medium of HMCLs significantly stimulated vessel formation compared with control or vascular endothelial growth factor (VEGF) treatment. The presence of anti-Tie2 blocking antibody completely blunted the proangiogenic effect of XG-6. Finally, our in vitro results were supported by the in vivo finding of Ang-1, but not Ang-2, mRNA and protein expression in purified MM cells obtained from approximately 47% of patients and by high BM angiogenesis in patients with MM positive for Ang-1, suggesting that the angiopoietin system could be involved, at least in part, in MM-induced angiogenesis.

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Decreased levels of sRAGE in follicular fluid from patients with PCOS

Reproduction ◽

10.1530/rep-16-0359 ◽

2017 ◽

Vol 153 (3) ◽

pp. 285-292 ◽

Cited By ~ 6

Author(s):

BiJun Wang ◽

Jing Li ◽

QingLing Yang ◽

FuLi Zhang ◽

MengMeng Hao ◽

...

Keyword(s):

Regression Analysis ◽

Granulosa Cells ◽

Follicular Fluid ◽

Cultured Cells ◽

Dependent Manner ◽

Vascular Endothelial ◽

Protein Levels ◽

Pcos Group ◽

Mrna And Protein Expression ◽

Endothelial Growth Factor

This study aimed to explore the association between soluble receptor for advanced glycation end products (sRAGE) levels in follicular fluid and the number of oocytes retrieved and to evaluate the effect of sRAGE on vascular endothelial growth factor (VEGF) in granulosa cells in patients with polycystic ovarian syndrome (PCOS). Two sets of experiments were performed in this study. In part one, sRAGE and VEGF protein levels in follicular fluid samples from 39 patients with PCOS and 35 non-PCOS patients were measured by ELISA. In part two, ovarian granulosa cells were isolated from an additional 10 patients with PCOS and cultured. VEGF and SP1 mRNA and protein levels, as well as pAKT levels, were detected by real-time PCR and Western blotting after cultured cells were treated with different concentrations of sRAGE. Compared with the non-PCOS patients, patients with PCOS had lower sRAGE levels in follicular fluid. Multi-adjusted regression analysis showed that high sRAGE levels in follicular fluid predicted a lower Gn dose, more oocytes retrieved, and a better IVF outcome in the non-PCOS group. Logistic regression analysis showed that higher sRAGE levels predicted favorably IVF outcomes in the non-PCOS group. Multi-adjusted regression analysis also showed that high sRAGE levels in follicular fluid predicted a lower Gn dose in the PCOS group. Treating granulosa cells isolated from patients with PCOS with recombinant sRAGE decreased VEGF and SP1 mRNA and protein expression and pAKT levels in a dose-dependent manner.

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Tumour vascular endothelial growth factor (VEGF) mRNA in relation to serum VEGF protein levels and tumour progression in human renal cell carcinoma

Urological Research ◽

10.1007/s00240-003-0346-x ◽

2003 ◽

Vol 31 (5) ◽

pp. 335-340 ◽

Cited By ~ 29

Author(s):

B�rje Ljungberg ◽

Jan Jacobsen ◽

Stina H�ggstr�m-Rudolfssson ◽

Torgny Rasmuson ◽

Gudrun Lindh ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Tumour Progression ◽

Vascular Endothelial ◽

Vegf Mrna ◽

Human Renal Cell Carcinoma ◽

Protein Levels ◽

Endothelial Growth Factor ◽

Vegf Protein

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Yangjing capsule attenuates cyclophosphamide-induced deficiency of testicular microcirculation in mice

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v19i3.21 ◽

2020 ◽

Vol 19 (3) ◽

pp. 603-608

Author(s):

Baofang Jin ◽

Weihang Dong ◽

Dalin Sun ◽

Bin Cai ◽

Weimin Deng ◽

...

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Western Blot ◽

Microvascular Density ◽

Vascular Endothelial ◽

Protective Effects ◽

Protein Levels ◽

Mrna And Protein Expression ◽

High Concentrations ◽

Endothelial Growth Factor

Purpose: To explore the protective effects of Yangjing capsule (YC) on testicular microcirculation in a mouse model of deficiency of testicular microcirculation. Methods: Immunohistochemistry was applied to determine the effects of YC on microvascular density of mice. The protein level of CD34 and vascular endothelial growth factor A (VEGF A) was measured by western blot. The viability of Testicular cell line (TM4 cells) was examined by CCK-8 assay. Results: Histopathological changes demonstrated that CP-induced decrease of microvascular density of the mice was rescued by YC dose-dependently (p < 0.5). Western blot data showed that the protein levels of CD34 and VEGF A in CP group were significantly decreased, but dose-dependently increased by YC, respectively, following co-administration of CP + YC, compared with those in CP group (p < 0.5). The results from CCK-8 assay showed that the cell viability of TM4 cells increased with the amount of YC administered, and that high concentrations of YC (0.1 and 1 mg/mL) showed significant effects (p < 0.5). Moreover, YC showed little effect on VEGF A mRNA and protein expression in TM4 cells. Conclusion: YC may be considered an alternative therapeutic agent for the management of testicular microcirculation disease. However, further studies are required to ascertain this. Keywords: Yangjing Capsule, Testicular microcirculation, Cyclophosphamide, Vascular endothelial growth factor A

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Effects of high glucose on vascular endothelial growth factor expression in vascular smooth muscle cells

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1997.273.5.h2224 ◽

1997 ◽

Vol 273 (5) ◽

pp. H2224-H2231 ◽

Cited By ~ 23

Author(s):

Rama Natarajan ◽

Wei Bai ◽

Linda Lanting ◽

Noe Gonzales ◽

Jerry Nadler

Keyword(s):

Vascular Endothelial Growth Factor ◽

Smooth Muscle ◽

Growth Factor ◽

Vascular Smooth Muscle Cells ◽

Vascular Endothelial ◽

Ang Ii ◽

Vegf Expression ◽

Vegf Mrna ◽

Mrna And Protein Expression ◽

Endothelial Growth Factor

Vascular endothelial growth factor (VEGF), in addition to its growth-promoting effects on endothelial cells, can also increase vascular permeability and monocyte migration. It has therefore been implicated in the pathogenic neovascularization associated with diabetic retinopathy and atherosclerosis. However, the factors regulating VEGF expression in the vascular wall are not fully understood. In this study, we examined the regulation of VEGF expression in vascular smooth muscle cells (VSMC) by hyperglycemia as well as by angiotensin II (ANG II). We also examined whether the 12-lipoxygenase (12-LO) product 12-hydroxyeicosatetraenoic acid (12-HETE) can alter VEGF expression, since 12-LO products of arachidonic acid have angiogenic properties, and ANG II as well as high glucose (HG, 25 mM) can increase 12-LO activity and expression in VSMC. Studies were carried out in human (HSMC) or porcine VSMC (PSMC), which were cultured for at least two passages under normal glucose (NG, 5.5 mM) or HG conditions. HG culture alone increased the expression of VEGF mRNA and protein in both HSMC and PSMC. Furthermore, ANG II treatment significantly induced VEGF mRNA and protein expression only in VSMC cultured in HG and not NG. In addition, 12-HETE significantly increased VEGF mRNA and protein expression in HSMC cultured in NG as well as in HG. Cells cultured in HG also secreted significantly greater amounts of VEGF into the culture medium. These results suggest that elevated VEGF production under HG conditions may play a role in the accelerated vascular disease observed in diabetes.

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Prolonged hypoxia increases vascular endothelial growth factor mRNA and protein in adult mouse brain

Journal of Applied Physiology ◽

10.1152/jappl.1999.86.1.260 ◽

1999 ◽

Vol 86 (1) ◽

pp. 260-264 ◽

Cited By ~ 63

Author(s):

Ning-Tsu Kuo ◽

David Benhayon ◽

Ronald J. Przybylski ◽

Richard J. Martin ◽

Joseph C. LaManna

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Protein Expression ◽

Feedback System ◽

Vascular Endothelial ◽

Vegf Mrna ◽

Protein Levels ◽

Adult Mice ◽

Endothelial Growth Factor ◽

The Brain

Brain hypoxia induces an increase in brain vascularity, presumably mediated by vascular endothelial growth factor (VEGF), but it is unclear whether VEGF is required to maintain the increase. In these studies, brain VEGF mRNA and protein levels were measured in adult mice kept in hypobaric chambers at 0.5 atm for 0, 0.5, 1, 2, 4, 7, and 21 days. Hypoxia was accompanied by a transient increase of VEGF mRNA expression: twofold by 0.5 day and a maximum of fivefold by 2 days; these were followed by a decrease at 4 days and a return to basal levels by 7–21 days. VEGF protein expression induced by hypoxia was bimodal, initially paralleling VEGF mRNA. There was an initial small increase at 12 h that reached a maximum by day 2, and, after a transient decrease on day 4, the protein expression increased again on day 7 before it returned to normoxic levels after 21 days. Thus, despite continued hypoxia, both VEGF mRNA and protein levels returned to basal after 7 days. These data suggest a metabolic negative-feedback system for VEGF expression during prolonged hypoxia in the brain.

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Experimental Methods to Simulate and Evaluate Postsurgical Peripheral Nerve Scarring

Journal of Clinical Medicine ◽

10.3390/jcm10081613 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1613

Author(s):

Alessandro Crosio ◽

Giulia Ronchi ◽

Benedetta Elena Fornasari ◽

Simonetta Odella ◽

Stefania Raimondo ◽

...

Keyword(s):

Peripheral Nerve ◽

Peripheral Nerves ◽

Experimental Methods ◽

Scar Tissue ◽

Experimental Models ◽

Tissue Formation ◽

Surgical Interventions ◽

Pubmed Database ◽

Different Types ◽

Scar Tissue Formation

As a consequence of trauma or surgical interventions on peripheral nerves, scar tissue can form, interfering with the capacity of the nerve to regenerate properly. Scar tissue may also lead to traction neuropathies, with functional dysfunction and pain for the patient. The search for effective antiadhesion products to prevent scar tissue formation has, therefore, become an important clinical challenge. In this review, we perform extensive research on the PubMed database, retrieving experimental papers on the prevention of peripheral nerve scarring. Different parameters have been considered and discussed, including the animal and nerve models used and the experimental methods employed to simulate and evaluate scar formation. An overview of the different types of antiadhesion devices and strategies investigated in experimental models is also provided. To successfully evaluate the efficacy of new antiscarring agents, it is necessary to have reliable animal models mimicking the complications of peripheral nerve scarring and also standard and quantitative parameters to evaluate perineural scars. So far, there are no standardized methods used in experimental research, and it is, therefore, difficult to compare the results of the different antiadhesion devices.

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