Therapeutic potential of traditional Chinese medicine for vascular endothelial growth factor / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Vascular endothelial growth factor (VEGF) is the main regulator of physiological angiogenesis during embryonic development, bone growth, and reproductive function, and it also participates in a series of pathological changes. Traditional Chinese medicine (TCM), with a history of more than 2000 years, has been widely used in clinical practice, while the exploration of its mechanisms has only begun. This review summarizes the research of recent years on the influence of TCM on VEGF. It is found that many Chinese medicines and recipes have a regulatory effect on VEGF, indicating that Chinese medicine has broad prospects as a complementary and alternative therapy, providing new treatment ideas for clinical applications and the theoretical basis for research on the mechanisms of TCM.

Expresión de VEGF-A y VEGFR2 en Miocardio de Pollos Tratados con Ácido Acetilsalicílico (AAS) / VEGF-A and VEGFR2 Expression in Chicken Myocardium Treated with Acetylsalicylic Acid (AAS)

RESUMEN: Los niveles de VEGF y su unión a sus receptores son etapas claves en la regulación de la angiogénesis. El ácido acetilsalicílico (AAS), ampliamente utilizado en tratamiento post infarto al miocardio ha mostrado poseer un efecto antiangiogénico en modelos tumorales. Este efecto potencialmente contraproducente requiere ser estudiado en miocardio. El objetivo del presente trabajo es cuantificar el efecto de AAS y de ácido salicílico (AS) sobre la vascularización en membrana alantocoriónica (MAC) y sobre los niveles de VEGF-A y VEGFR2 en miocardio de embriones de pollo. Para ello, treinta fetos de pollo White Leghorn fueron instilados a los 10 días de gestación con 60 µL de DMSO 0,1 % (control) o conteniendo además 0,3 µmol de AAS o AS. A las 48 horas se realizó procesamiento histológico de MAC para recuento de vasos sanguíneos y de tejido cardíaco para cuantificar VEGF-A y VEGFR2 por inmunohistoquímica. La inmunorreactividad fue cuantificada mediante Image J. Tanto AAS como AS disminuyeron la densidad microvascular de MAC. En miocardio, AAS aunque no AS, disminuyó la concentración de VEGFR2. No hubo efecto sobre VEGF-A. En nuestro modelo experimental, fetos de pollo a los 10 días de gestación también se observó el efecto inhibidor de AAS sobre la angiogénesis en MAC. La disminución de VEGFR2 en cardiomiocitos sugiere que AAS también afecta la angiogénesis en miocardio sano, modificando la disponibilidad del receptor a VEGF. Estos hallazgos nos permiten postular que AAS podría interferir con la regeneración de tejido, en situaciones como post infarto al miocardio.

SUMMARY: The VEGF levels and its binding to its receptors are key stages in the regulation of angiogenesis. Acetylsalicylic acid (ASA), widely used in post-myocardial infarction treatment, has been shown to have an anti-angiogenic effect in tumor models. This potentially counterproductive effect requires to be studied in myocardium. The aim of this study is to quantify the effect of ASA and salicylic acid (SA) on the vascularization in chick allantochorionic membrane (CAM) and on the levels of VEGF-A and VEGFR2 in myocardium of chicken embryos. Thirty White Leghorn chicken fetuses were instilled at 10 days of gestation with 60 mL of 0.1 % DMSO (control) or also containing 0.3 mmol of ASA or SA. After 48 hours, CAM histological processing was performed to count blood vessels and heart tissue to quantify VEGFA and VEGFR2 by immunohistochemistry. Immunoreactivity was quantified by Image J. Both ASA and SA decreased CAM microvascular density. In myocardium, AAS, although not SA, decreased the concentration of VEGFR2. There was no effect on VEGF-A. In our experimental model, chicken fetuses at 10 days of gestation, the inhibitory effect of ASA on angiogenesis in CAM were also observed. The decrease in VEGFR2 in cardiomyocytes suggests that ASA also affects angiogenesis in healthy myocardium, modifying the availability of the receptor to VEGF. These findings allow us to postulate that ASA could interfere with tissue regeneration, when it is required, as post myocardial infarction.

Pien Tze Huang promotes HepG2 apoptosis via regulating ANXA1/VEGF signaling pathway / 中国药理学通报

Aim To explore the effect of Pien Tze Huang (PZH) on inhibiting HepG2 cells via regulating ANXA1/VEGF signaling pathway and the underlying mechanism. Methods HepG2 cells were treated with different concentrations (1, 1 0, 100 mg • L ~ l ) of PZH. MTT assay and colony formation assay were used to calculate cell viability and cell survival. Western blot was used to determine the expression of Bax, Bcl-2, cleaved caspase-3, cleaved caspase-9, and ANXA1/VEGF signaling pathway protein, such as ANXA1, VEGF, VEGFR, NF-KB P 5 0 . Results Compared with normal group, different concentrations of PZH inhibited HepG2 cells in a dose-dependent manner, inhibited the colony formation, promoted the expression of apoptotic expression, promoted the expression of ANXA1 protein, inhibited the expression of VEGF, VEGFR, and N F - K B P 5 0 as well. Conclusions PZH can inhibit the activity of HepG2 cells in vitro. Its main mechanism is related to the promotion of apoptotic protein in HepG2 cells, the promotion of cell ANXA1 protein, and the inhibition of VEGF/VEGF receptor and NF-KB signaling pathway.

Research advance of retinal neovascularization inhibitory factor / 国际眼科杂志(Guoji Yanke Zazhi)

The normal growth of blood vessels is the result of dynamic balance of angiogenic factor and inhibitory factor in vascular tissue.However, when the balance is broken, the growth of new blood vessels will be induced.Endogenous angiogenesis inhibitory factor, is a group of negative feedback molecules produced by the body itself that inhibit angiogenesis.Its function of inhibiting angiogenesis is mainly realized by promoting the binding of angiogenic factor to its receptor, or its downstream angiogenesis signal, or promoting vascular endothelial apoptosis.The study of angiogenesis inhibitory factor has potential clinical significance for the prevention and treatment of retinal neovascularization.Recent studies on retinal neovascularization inhibitory factor are reviewed in this paper.

VEGF-VEGFR Signals in Health and Disease

Vascular endothelial growth factor (VEGF)-VEGF receptor (VEGFR) system has been shown to play central roles not only in physiological angiogenesis, but also in pathological angiogenesis in diseases such as cancer. Based on these findings, a variety of anti-angiogenic drugs, including anti-VEGF antibodies and VEGFR/multi-receptor kinase inhibitors have been developed and approved for the clinical use. While the clinical efficacy of these drugs has been clearly demonstrated in cancer patients, they have not been shown to be effective in curing cancer, suggesting that further improvement in their design is necessary. Abnormal expression of an endogenous VEGF-inhibitor sFlt-1 has been shown to be involved in a variety of diseases, such as preeclampsia and aged macular degeneration. In addition, various factors modulating angiogenic processes have been recently isolated. Given this complexity then, extensive studies on the interrelationship between VEGF signals and other angiogenesis-regulatory systems will be important for developing future strategies to suppress diseases with an angiogenic component.

VEGF-VEGFR Signals in Health and Disease

Vascular endothelial growth factor (VEGF)-VEGF receptor (VEGFR) system has been shown to play central roles not only in physiological angiogenesis, but also in pathological angiogenesis in diseases such as cancer. Based on these findings, a variety of anti-angiogenic drugs, including anti-VEGF antibodies and VEGFR/multi-receptor kinase inhibitors have been developed and approved for the clinical use. While the clinical efficacy of these drugs has been clearly demonstrated in cancer patients, they have not been shown to be effective in curing cancer, suggesting that further improvement in their design is necessary. Abnormal expression of an endogenous VEGF-inhibitor sFlt-1 has been shown to be involved in a variety of diseases, such as preeclampsia and aged macular degeneration. In addition, various factors modulating angiogenic processes have been recently isolated. Given this complexity then, extensive studies on the interrelationship between VEGF signals and other angiogenesis-regulatory systems will be important for developing future strategies to suppress diseases with an angiogenic component.

Influence of Yifeiqinghua Granules on Expression of Vascular Endothelial Growth Factors and Its Receptor KDR in Lewis Lung Cancer Mice / 中国中医药信息杂志

Objective To clarify the mechanism of Yifeiqinghua granules on tumor angiogenesis and tumor inhibition by observing its influence on the expression of vascular endothelial growth factor (VEGF) and its receptor KDR in Lewis lung cancer mice. Methods Eighty C57BL/6 inbred mice subcutaneously planted with Lewis tumor cell suspension 0.2 mL were randomly divided into 8 groups:blank group, control group, low-, medium- and high-dose of Yifeiqinghua granules groups, Gefitinib group, Gefitinib plus medium-dose of Yifeiqinghua granules group and CTX group, 10 mice in each group. Drugs were administrated from the second day. CTX group was administered on third day and seventh day, other groups were administered for 14 days. The mice were killed to take the tumor on the 15th days. VEGF and KDR expression in tumor tissue was measured by Western Blot. Results Compared with the blank group, the expression of VEGF was significantly decreased in the low- and medium-dose of Yifeiqinghua granules groups (P<0.01). Gefitinib group’s VEGF value was lower than the blank group (P<0.05). Compared with the blank group, the expression of VEGF was significantly decreased in the high-dose of Yifeiqinghua granular group, Gefitinib plus medium-dose of Yifeiqinghua granules group and CTX group (P<0.001). Compared with the blank group, the expression of KDR was significantly decreased in the low-, medium- and high-dose of Yifeiqinghua granules groups, Gefitinib group and CTX group (P<0.05). Conclusions The possible mechanism of Yifeiqinghua granules in treating non-small cell lung cancer is reducing the expression of VEGF and KDR, thus play an inhibitory effect.

The Effect of Antipsychotic Drug Treatment on Serum VEGF, sVEGFR-1, and sVEGFR-2 Level in Schizophrenia: A Preliminary Study

OBJECTIVES: Vascular endothelial growth factor(VEGF), one of potent cytokines, and its receptors were related with various biological functions and pathological conditions. The purpose of this study was to investigate the changes of serum level of free VEGF, soluble VEGFR-1, and soluble VEGFR-2 after treatment with atypical antipsychotic drug in schizophrenia. METHOD: The schizophrenic patients were diagnosed with DSM-IV and were prospectively followed up for 4 and 8 weeks. Thirteen schizophrenic patients were evaluated their clinical assessment with serum levels of free VEGF, sVEGFR-1, sVEGFR-2, and positive and negative symptom scale(PANSS) at baseline, 4 weeks, and 8 weeks after treatment with atypical antipsychotic drug. Thirteen normal control subjects were recruited and matched with the patient group by age and sex. RESULT: The serum level of free VEGF(295.2+/-43.7pg/ml)and sVEGFR-2(8259+/-336.7) at baseline(before treatment) in schizophrenic patients were not significantly different, compared with the control group(199.0+/-28.8 and 8481+/-371.9) respectively. However, the serum level of sVEGFR-1(86.2+/-10.3, p<0.05) was significantly increased in the schizophrenic patients compared with the control group(59.0+/-6.4). After treatment with antipsychotic drug, the serum levels of free VEGF at 4 weeks(338.9+/-56.5) and 8 weeks(309.5+/-58.7) were not significantly, different compared with baseline. But the serum levels of sVEGFR-1 was significantly decreased at 8 weeks(57.3+/-6.3, p<0.05) after antipsychotic drug treatment. The serum levels of sVEGFR-2 were decreased at 4 weeks(7761+/-403.0, p<0.05) and 8 weeks(7435+/-333.5, p<0.05) compared with baseline. CONCLUSION: The decreased serum level of sVEGFR-1 and sVEGFR-2 might be affected by dopaminergic system which was influenced by antipsychotic drug.

The soluble receptor flt-1_(n3) of vascular endothelial growth factor inhibits bladder carcinoma angiogenesis and growth / 中华泌尿外科杂志

Objective To verify whether the extracellular domain of flt 1 has anti angionesis activity in vivo. Methods A recombinant plasmid pcDNA 3.1/flt 1 n3 was transfected into the human bladder carcinoma EJ cell by lipofectamin, which was constructed by inserting the N terminal first three IgG like domain of VEGF receptor flt 1 (flt 1 n3 ) into eukaryotic expression vector pcDNA 3.1. Results The rflt 1 was functionally expressed in stably transfected EJ and the product secreted in the medium could be specifically bind to rhVEGF165.The result showed that the bladder cancer transfected with rflt 1 had a lower microvessle density than the control. Conclusions It is proved that the expressed rflt 1 n3 can inhibit tumor growth and angiogenesis in nude mice model.

The Influence of Polycationic Polypetride on the Transfective Efficacy of NonViral Vector / 中国肿瘤生物治疗杂志

Objective: Purpose to investigate the different in vitro function of targetable non-viral vector containing poly-L-lysine or protamine. Methods: Using GV1 and GV2 targetable non-viral vectors, the influences of the poly-L-lysine and protamine on in vitro gene transfer efficiency and the course of gene expression were observed. Results: ?-galactosidase was expressed at intermediate level (50% ) in A375 cells using a complex containing either protamine or poly-L-lysine. Howerver, in case of ABAE cells, ?－galactosidase expression level was low (20% ) transferred with a comPlex containing protamine. On the contrary, ?－ galactosidase expression was at high level (70% ) provided that protamine was replaced with poly-L-lysine. In addition, ?-galac- tosidase activity reached the peak at the 6th day after transfection with the complex containing protamine. The expression was not altered with subsequent subcultures, at least for 3 passages. Using poly-L-lysine, the expression peak in A375 reached the peak at the 7th day after transfection, but the level declined along with subsequent passages of cells. Conclusion: The apllication of protamine in VEGF receptor mediated gene delivery system was limited.

Research on function of VEGF receptor 3 high-bound peptide / 第三军医大学学报

Objective To assess the function of the peptide (WHGSLKQNLWWY) through measuring the a functional affinity constant between humanized VEGF receptor 3 and peptide, then comparing it to that between VEGF D and peptide. Methods After coating the peptide by BSA binding with glutaral couple, the best concentration, best time of peptide to coat the plate and the coating coefficient were determined. With cognizance of the advantages of solid phase method in time consumption and convenience, we assessed the functional affinity constant of engineered peptide and VEGF-D with non-competitive ELISA method. We plotted the D(410) standard curve of the binding reaction of peptide and VEGFR3/Fc using four grades of concentration of VEGFR3/Fc and a series of consistency of peptide. We got the consistency of peptide at the half of maximal OD value through the standard curve and calculated the affinity constant by law of Mass Action. Results The affinity constant of peptide was between 10 7-10 8 mol/L -1, which was only smaller by 10 times than that of VEGF-D. Conclusion The peptide we got from the peptide liberary can strongly bind to the target (VEGFR3), thus providing a theoretical basis for its pharmacal use.