Vascular endothelial growth factor (VEGF) exerts its biological functions by its specific VEGF receptors (VEGFRs), which includes VEGFR-1, VEGFR-2, VEGFR-3, neuropilin-1 and neuropilin-2. These VEGF receptors not only distribute in endothelial cells, but also in epidermal keratinocytes. VEGFRs may play a significant role in pathogenesis of the epidermal neoplasm and the VEGF-VEGFR signaling pathway may be a novel therapy target for neoplasm derived from epidermis.
Animals ; Epidermis ; metabolism ; Humans ; Neoplasms ; metabolism ; Neuropilins ; genetics ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-3 ; genetics ; metabolism

OBJECTIVETo observe the expression levels of hippocampal vascular endothelial growth factor (VEGF), fms-like tyrosine kinase-1 (flt-1), basic fibroblast growth factor (bFGF), and basic fibroblast growth factor receptor (bFGF-r) in vascular dementia (VD) rats, thus studying the angiogenesis mechanism of moxibustion in VD.

METHODSSixty male elderly Wistar rats were selected. The VD rat model was prepared by bilateral carotid artery occlusion and reperfusion of sodium nitroprusside injection. The model rats were divided into 3 groups by the random digit table, i. e., the moxibustion group, the Western medicine group, and the model group. A sham-operation control group was also set up. In the moxibustion group rats was acupunctured at Baihui (GV20), Shenting (GV14), and Dazhui (GV24). Aniracetam was given to rats in the Western medicine group by gastrogavage for 2 therapeutic courses, 15 days as one course. The learning and memory results were observed by the neuroethological score in combination of step-down avoidance test before treatment and by the end of the 2nd course respectively. The expression levels of hippocampal VEGF, flt-1, bFGF, and bFGF-r of all rats were detected using immunohistochemical assay.

RESULTSAfter 2 courses of treatment, statistical difference existed in the latent period, the error times, and the neuroethological score in the moxibustion group and the Western medicine group when compared with the model group (P < 0.01, P < 0.05). Statistical difference existed in the latent period and the neuroethological score between the moxibustion group and the Western medicine group (P < 0.05), which indicated that moxibustion and Western medicine showed significant effects in improving the latent period, decreasing the error times and the neuroethological score. Better results were obtained in the moxibustion group than in the Western medicine group (P < 0.01, P < 0.05). Statistical difference of the average grey level (AGL) of hippocampal VEGF, flt-1, and bFGF existed in the moxibustion group and the Western medicine group when compared with the model group. Statistical difference of the bFGF-r expression existed only between the moxibustion group and the model group. Statistical difference of the VEGF and flt-1 expressions existed between the moxibustion group and the Western medicine group (P < 0.05).

CONCLUSIONSMoxibustion showed confirmative effects in improving the behavioral score and memory performance in VD rats. Its mechanisms might lie in that moxibustion regulated and controlled the expression levels of hippocampal VEGF, flt-1, bFGF, and bFGF-r in VD rats. Particularly it up-regulated the expression levels of key factors VEGF and flt-1, promoted the angiogenesis in the vital parts, and ultimately stimulated the repairing mechanisms of cerebral nerve injury.

Animals ; Dementia, Vascular ; metabolism ; therapy ; Fibroblast Growth Factor 2 ; metabolism ; Hippocampus ; metabolism ; Male ; Moxibustion ; Rats ; Rats, Wistar ; Receptor, Fibroblast Growth Factor, Type 2 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism

OBJECTIVETo investigate the effect of transfection with human soluble vascular endothelial growth factor receptor-1(sFlt-1) gene on cell growth and vascular endothelial growth factor (VEGF) concentration of the culture supernatant in human colon cancer cell line Lovo.

METHODSThe recombinant plasmid pcDNA3-sFlt-1 containing sFlt-1 gene was transfected into Lovo cells by Lipofectamine 2000, which was identified by RT-PCR and ELISA. The effect of sFlt-1 protein on cell growth and VEGF expression in Lovo cells were investigated by MTT and ELISA.

RESULTSThe recombinant plasmid pcDNA3-sFlt-1 was successfully transfected into Lovo cells. The sFlt-1 expression was identified by RT-PCR and ELISA, which inhibited the growth of Lovo cells and reduced the VEGF concentration in the culture supernatant compared with control. The inhibitory rates of proliferation of Lovo cells via MTT assay after 2,14,21 and 28 days were(23.92+/-9.16)%, (13.98+/-10.21)%,(22.54+/-11.92)% and (33.43+/-9.34)% respectively. Compared with the control groups, the differences were significant (P<0.05, P<0.01).

CONCLUSIONTransfection with sFlt-1 gene into Lovo cells results in the expression of sFlt-1 protein, which possesses high biological activity and inhibits the growth of cancer cells.

Cell Line, Tumor ; Genetic Vectors ; Humans ; Transfection ; Vascular Endothelial Growth Factor Receptor-1 ; genetics ; metabolism

OBJECTIVETo study the expression of vascular endothelial growth factor (VEGF) and its receptors, the fms-like tyrosine-1 (flt-1) and kinase insert domain-containing receptor (KDR) in endometrial carcinoma and investigate the functions of VEGF and its receptors for endometrial carcinoma angiogenesis and its relation to the grade of tumor.

METHODSImmunocytochemistry and in situ hybridization technique were used to measure the level of VEGF, flt-1, KDR protein and mRNA in endometrial carcinoma tissue from 23 patients and endometrial samples from 6 normal menopausal women. A few endometrial carcinoma samples were homogenized for Western blot analysis. The blood vessel density was estimated by counting blood vessels stained with endothelial marker VIII factor.

RESULTSThe VEGF and its receptors were widely expressed in the cytoplasm of endothelial cells and tumor cells of endometrial carcinoma. The level of VEGF protein in endothelial cells and endometrial cancer cells of grade II and III tumor tissues was higher than that in grade I and normal menopausal endometrium (P < 0.05). VEGF mRNA did not show higher expression with the increase of tumor grade but its expression in normal tissue was lower than that in cancer (P < 0.05). The expression of flt-1 protein and mRNA in endothelial cells got higher in III than in grade II and I (P < 0.05), but invariable in cancer cells (P > 0.05), flt-1 expression in cancer was higher than that in normal menopausal endometrium either in endothelial cells or in cancer cells (P < 0.05). The expression of KDR protein in endothelial and cancer cell was high but did not alter with the increase of tumor grade (P > 0.05), the level of its mRNA was higher in cancer than that in normal tissue (P < 0.05). The microvascular density in grade III (48 +/- 12) was higher than that in grade II (26 +/- 16), grade I (27 +/- 14) and normal menopausal tissue (26 +/- 11, P < 0.05).

CONCLUSIONSThe expression pattern of VEGF, flt-1 and KDR protein and mRNA increased with the increase of tumor grade in endometrial carcinoma indicates that VEGF and its receptors contribute to the neovascularization of tumors and is one of the factors that relate to rapid tumor growth of endometrial carcinoma.

Endometrial Neoplasms ; metabolism ; physiopathology ; Endothelial Growth Factors ; genetics ; metabolism ; Extracellular Matrix Proteins ; metabolism ; Female ; Gene Expression ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Lymphokines ; genetics ; metabolism ; Neovascularization, Pathologic ; RNA, Messenger ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism ; Vascular Endothelial Growth Factors

OBJECTIVETo observe the effect of Panax notoginseng (PN) on pathological features in chronic subdural hematoma (CSDH) rabbits and its mechanisms.

METHODSA stable pathological animal model similar to CSDH in humans could be established using subdural injections of small number of blood through a subdural pre-catheter in rabbits. After successful modeling, 18 rabbits were randomly divided into the model group, the low dose PN group (0.125 g/kg), and the high dose PN group (0.250 g/kg), 6 in each group. Normal saline was given to rabbits in the model group, while PN power was given to those in the PN groups by gastrogavage for 6 successive days. Pathologic features of the hematoma outer membrane were observed by HE staining. The activity of SOD and the content of MDA in the hematoma outer membrane were examined by the colorimetric method. Expressions of CD31, CD34, and VEGF in the hematoma outer membrane were observed by immunohistochemical assay. Expressions of VEGF in the peripheral blood and the subdural hematoma were detected by enzyme-linked immunosorbent assay (ELISA). Expressions of VEGFR-1 and VEGFR-2 in the hematoma outer membrane were detected by Western blot.

RESULTSCompared with the model group, the inflammatory reaction was comparatively lessen and the proliferation of the fibrous tissue was relatively mature in the low and high dose PN groups. The activity of SOD increased (P < 0.05); expressions of CD31 and CD34 were reduced (P < 0.01); VEGF expression in the residual hematoma fluid decreased (P < 0.05) in the high dose PN group. Expressions of VEGF and VEGFR-2 were all reduced in the high and low dose PN groups (P < 0. 05, P < 0.01). Compared with the low dose PN group, expressions of CD31 and CD34 were reduced (P < 0.01), and the VEGFR-2 expression was also reduced (P < 0.05) in the high dose PN group.

CONCLUSIONSPN could promote the fibrous repairing of subdural hematoma in CSDH rabbits. It also lessened inflammation and oxidative injury of the hematoma outer membrane and reduced expressions of VEGF. The pathological angiogenesis could be reduced through influencing VEGFR-2 receptor pathways, which might be an important mechanism.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Hematoma, Subdural, Chronic ; metabolism ; pathology ; Panax notoginseng ; chemistry ; Rabbits ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

OBJECTIVETo study the expressions of the vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) in the testis, epididymis and epididymal sperm of adolescent rats and explore the functions of both the proteins in the male reproductive system.

METHODSThe expressions of VEGF and Flt-1 were detected in 20 adolescent SD rats, immunohistochemical staining used for both the testis and the epididymis and immunofluorescent staining for sperm.

RESULTSVEGF and Flt-1 proteins were specifically present in the testis, epididymis and sperm. In the testis, VEGF immunoreactive particles were localized in the cytoplasm of spermatogenic cells, the developing acrosome of spermatids, Sertoli cells and Leydig cells, while Flt-1 expressed mainly in the developing acrosome of spermatids and Leydig cells. In the epididymis, the cell-specific and region-specific expressions of VEGF and Flt-1 proteins were observed in the principal cells of epididymal epithelia, VEGF in the whole epididymis, while Flt-1 only in the caput and cauda segments. Both VEGF and Flt-1 were localized in the acrosome of the sperm head as well as in the neck, middle and principal segments of the sperm tail.

CONCLUSIONThe specific expression patterns of VEGF and Flt-1 in the rat testis, epididymis and sperm indicate that they may independently or collectively affect spermatogenesis and spermiotelcosis in either an autocrinological or a

Animals ; Epididymis ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Sexual Maturation ; Spermatozoa ; metabolism ; Testis ; metabolism ; Vascular Endothelial Growth Factor A ; biosynthesis ; Vascular Endothelial Growth Factor Receptor-1 ; biosynthesis

OBJECTIVETo explore the role of angiogenesis in bone marrow in acute myeloid leukemia (AML).

METHODSBone marrow culture supernatant was assayed for vascular endothelial growth factor (VEGF) by ELISA, bone marrow biopsies from 28 newly diagnosed AML patients were assayed for microvessel density (MVD), VEGF and its receptors KDR, Flt-1 by immunohistochemical staining before and after induction chemotherapy.

RESULTSCulture supernatant of AML bone marrow mononuclear cells showed higher amount of VEGF (425.31 ng/L) than that of control (140.12 ng/L). The VEGF and KDR expressions and MVD were significantly higher in newly diagnosed AML patients (78.6%, 78.6% and 7.1%, respectively) than that of control group (P < 0.05). There was a positive correlation between VEGF, KDR and MVD. The positive rate of VEGF, KDR and MVD reduced to normal after the patients achieved complete remission, while in non-remission patients did not. Kaplan-Meier analysis showed that the survival time was longer in VEGF negative group than in VEGF positive group. The pre-treatment MVD and VEGF had no correlation with survival time.

CONCLUSIONSThere is remarkable angiogenesis in AML and VEGF/KDR signaling pathway takes an important role in the pathological angiogenesis. VEGF could be used as a prognostic factor in AML.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; therapy ; Male ; Middle Aged ; Neovascularization, Pathologic ; etiology ; Signal Transduction ; Vascular Endothelial Growth Factor A ; analysis ; physiology ; Vascular Endothelial Growth Factor Receptor-1 ; analysis ; Vascular Endothelial Growth Factor Receptor-2 ; analysis

OBJECTIVETo examine the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR1, VEGFR2) in transdifferentiated human proximal tubular epithelial (HK-2) cell induced by transforming growth factor beta1 (TGFbeta1).

METHODSThe transdifferentiation of HK-2 cells was detected by evaluation of expression of alpha-SMA by cytoimmunochemistry and RT-PCR. The VEGF mRNA was evaluated with RT-PCR. The secreted VEGF in the culture media was measured with ELISA. The cellular VEGF, VEGFR1, and VEGFR2 were measured with Western blot.

RESULTSThe immunostain of alpha-SMA were positive in HK-2 cell induced by TGFbeta1 at the concentration of 5 and 8 ng/ml for 72 h. The expression of alpha-SMA mRNA was induced by TGFbeta1 in concentration- and time-dependent manners. The expressions of mRNA and protein of VEGF were upregulated by TGFbeta1 at the concentration of 0.1 and 1 ng/ml for 72 h and at the concentration of 8 ng/ml for 12 h and 24 h when compared with the control. But expressions of mRNA and protein of VEGF were downregulated by TGFbeta1 at the concentration of 3, 5, and 8 ng/ml for 72 h and at the concentration of 8 ng/ml for 36, 48, and 72 h, respectively. Meanwhile, Protein levels of VEGFR1 and VEGFR2 were upregulated by TGFbeta1 in concentration- and time- dependent manners.

CONCLUSIONSIncreased expression of VEGFR1 and VEGFR2 and two-phase change in VEGF expression occurred in the process of tubular epithelial transdifferentiation induced by TGFbeta1. Reduced expression of VEGF may contribute to tubular epithelial transdifferentiation in a vicious circle.

Cell Differentiation ; Epithelial Cells ; cytology ; Humans ; Kidney Tubules, Proximal ; cytology ; RNA, Messenger ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1 ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

AIMTo investigate the effects of Angelica sinensis on the expression of Flt-1, Flk-1 mRNA after the ischemic brain injury in rats.

METHODSWistar rats randomly divided into two groups: group A rats underwent middle cerebral artery occlusion (MCAO) for 2 hours by suture, group B rats underwent MCAO for 2 hours meanwhile received treatment with Angelica sinensis (5g/kg). At 1 st d, 3 rd d and 7 th d after reperfusion, 36 rats( n = 18 in each group) were assessed by neurological scale and brain tissue was taken to assess the lesion ration with 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. The other rats (n = 3 at different time points in each group) were decapitated at 3 h, 6 h, 12 h , 1 st d, 3 rd d, 7 th d after reperfusion. Quantitative reverse transcription and polymerase chain reaction (RT-PCR) technique was used to examine the gene expression of Flt-1, Flk-1.

RESULTSThe neurologic deficit score of rats in group B decreased significantly compared with group A at the same time point (P < 0.05). The infarct volume of group A was significant greater than group B at the same time point after reperfusion (P < 0.01). The results of RT-PCR revealed that the gene expression of Flt-1, Flk-1 in the two groups increased from 3 h after reperfusion and reached its peak at the time of 3 rd d after reperfusion, then declined gradually. The gene expression of Flt-1, Flk-1 in the group B was significantly increased than group A at the same time point (P < 0.01). The gene expression of Flk-1 was positive correlated with Flt-1 in two groups (r = 0.957).

CONCLUSIONThe increasing amount of Flt-1, Flk-1 expression was enhanced by Angelica sinensis following transient interruption of cerebral blood flow in rats.

Angelica sinensis ; Animals ; Brain Ischemia ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

OBJECTIVEBone-marrow derived mesenchymal stem cells (BMSC) have the potential to differentiate into endothelial cells. The aim of the study was to investigate the induction process of BMSC by B16 melanoma cells in vitro and to analyze the role of VEGF-a in the process.

METHODSA co-culture system containing BMSC and B16 melanoma cells based on transwell indirect model was established, and the induction process of BMSC by B16 melanoma cells was studied in vitro.

RESULTSBMSC were isolated from the bone marrow of C57 mice. BMSC expressed CD105, CD90, CD73, CD44 and CD166, and acquired expressin of endothelial phenotype markers including VEGFR-1, VEGFR-2 and Factor VIII after co-culture with B16 melanoma cells for 48 hours. The expression level of VEGFR-2 would be double and Factor VIII threefold more by extending the co-culture time to 72 hours. In the co-culture system, B16 melanoma cells also up-regulated the expression of VEGF-a.

CONCLUSIONSVEGF-a plays a significant role in the differentiation of BMSC into cells of endothelial phenotype, therefore, is important to tumor angiogenesis.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Coculture Techniques ; Endothelial Cells ; cytology ; metabolism ; Factor VIII ; metabolism ; Male ; Melanoma, Experimental ; metabolism ; pathology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Pilot Projects ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism