Objective To investigate the protection on the intrahepatic cholangiocyte mediated by hypoxic preconditioning (HP) after liver transplantation and the role of vascular endothelial growth factor (VEGF). Methods The model of autologous liver transplantation was established, and the rats were divided into 3 groups: autologous liver transplantation group, hypoxic preconditioning before operation group (HP group) and sham operation group. At 6, 12, 24, 48 h after operation, blood samples were collected for examination of the serum total bilirubin (TBIL), direct bilirubin (DBIL) and alkaline phosphatase (ALP), and the expression of VEGF was detected by immunohistochemical method. The pathological changes of cholangiocytes were observed by light microscope. Results As compared with autologous liver transplantation group, the levels of seurm TBIL, DBIL and ALP in HP group were lower (P＜0.05), while the expression of VEGF in HP group was higher at the whole process (P＜0.05). The degrees of billiary epithelium damage and inflammatory infiltration in autologous liver transplantation group were more severe than those in HP group. Conclusion HP has protective effect on cholangiocytes after liver transplantation, in which VEGF may play an important role.
【Abstract】Objective To evaluate the status of vascular endothelial growth factor (VEGF) expression in breast carcinoma and benign disease and define the relationship with age,menopause, tumor size，clinical stage，distant metastasis and lymph node metastasis. Methods Seventy cases of invasive ductal breast carcinomas,30 benign breast diseases and 7 adjacent nonneoplastic specimens were assessed for VEGF protein expression by immunohistochemistry LSAB method. Results VEGF were expressed more frequently in breast cancer than in benign diseases.VEGF was significantly correlated with axillary lymph node metastasis and distant metastasis,whereas no statistical correlation with other factors. Conclusion VEGF status has certain value to make differential diagnosis between malignant and benign breast diseases and predict the possibilities of distant and lymph node metastasis.
ObjectiveTo observe the effect of Delta-like ligand 4 (Dll-4) on the pathological structure of retina in early diabetic rats (DM) and its relationship with vascular endothelial growth receptor-2 (VEGFR-2).MethodsA total of 70 male Sprague-Dawley rats were randomly divided into normal group and DM group, with 10 and 60 rats in each group, respectively. The rats of DM group was induced by intraperitoneal injection of streptozotocin to established DM model. The rats with blood glucose recovery and death were excluded, and the final 60 rats were included in the statistics. Rats in the normal group were injected with an equal volume of citric acid-sodium citrate buffer. Rats in the DM group were divided into DM 1 month (DM 1m) group, DM 2 months (DM 2m) group, DM 3 months (DM 3m) group and DM 3m + Anti group, DM 3m + phosphate buffer solution (PBS) group by random number table method, and 10 rats in each group. In the DM 3m+Anti group, 4 μl of anti-Dll-4 polyclonal antibody was injected into the vitreous cavity, and the antibody concentration was 0.25 mg/ml. The DM 3m+PBS group was intravitreally injected with an equal volume of PBS. Five days after the injection, the rats were sacrificed. Rats in the DM 3m group and the normal group were not treated, and were sacrificed 3 months after the model was established. The structure and microvascular changes of the retina were observed by hematoxylin-eosin staining, and the total thickness of the retina was measured. The expression of Dll-4 and VEGFR-2 in the retina was detected by immunohistochemistry and fluorescence quantitative polymerase chain reaction (PCR). One-way analysis of variance was used to compare the expression of Dll-4 and VEGFR-2 in the retina of each group. The least significant difference t test was used to compare the two groups.ResultsLight microscopy showed that the retinal ganglion cells layer in the DM 3m group were obviously edematous, the inner and outer nuclear layers were thinner, the number of cells was reduced, the arrangement was disordered, the edema of outer plexiform layer was obvious, and the microvessels were abnormally dilated. In the DM 3m+Anti group, the edema of outer plexiform layer was lessened than that of the DM 3m group, and the other layers were not significantly different from the DM 3m group. Compared with the normal group, the total retinal thickness of the DM 3m group, the DM 3m+Anti group and the DM 3m+PBS group increased (t=5.596, 3.290, 4.286; P=0.000, 0.008, 0.002). Immunohistochemical staining showed that a small amount of Dll4 was positively expressed in the retinal ganglion cell layer of the normal group; a small amount of VEGFR-2 was positively expressed in the ganglion cell layer and the inner and outer nuclear layers. The positive expression of Dll-4 and VEGFR-2 in retinal vascular endothelial cells of DM 3m group increased significantly. The expression of Dll-4 was significantly decreased in the retinal layers and vascular endothelial cells of DM 3m+Anti group, while the expression of VEGFR-2 was significantly increased. There was no significant difference between the positive expression of Dll4 and VEGFR-2 in the DM 3m+PBS group and the DM 3m group. The results of real-time PCR showed that the relative expression of Dll-4 and VEGFR-2 mRNA in the DM 3m group was significantly higher than that in the normal group (t=6.705, 20.871; P＜0.05). Compared with DM 3m group, the relative expression of Dll-4 mRNA in DM 3m+Anti group decreased, and the relative expression of VEGFR-2 mRNA increased (t=2.681, 3.639;P＜0.05). The relative expressions of Dll-4 and VEGFR-2 mRNA in the DM 3m+PBS group and DM 3m group were not statistically significant (t=0.513, 0.657; P＜0.05).ConclusionsThe expression of Dll-4 in retinal vascular endothelial cells is gradually increased during the early retinopathy of DM rats. The expression of Dll-4 is inhibited, the expression of VEGFR-2 is up-regulated, and the plexus edema is alleviated.
Objective To investigate the effect of caveolion-1 on the growth and proliferation in human breastcancer MCF7 cell in vitro and in vivo and approach its mechanisms. Methods The plasmid pCI-neo-cav-1 and its corres-ponding empty vector （pCI-neo） were transfected into MCF7 cell line. The expressions of caveolin-1 and vascular endoth-elial growth factor （VEGF） in transfectants were subsequently confirmed by Western blot analysis. A pair of monoclonal cell lines， MCF7/cav-1 and MCF7/vec， were chose for future studies. The colony formation ability of tumor cells was detected by anchorage-independent growth assay. Xenograft tumor models in nude mice were established. Immunohisto-chemistry was used to characterize Ki-67 and VEGF levels in the tumors tissues. Results Caveolin-1 expression was up-regulated in the MCF7/cav-1 cell as compared with the MCF7/vec cell （P＜0.01） and the MCF7 cell （P＜0.01）. Caveolin-1 overexpression markedly reduced the capacity of the cells to form colonies in soft agar （P＜0.01）. Compared with the MCF7/vec cell， VEGF protein expression reduced in the MCF7/cav-1 cell （P＜0.01）. In vivo experiments in nude mice， the overexpression of caveolin-1 resulted in significant growth inhibition of the xenograft breast tumors. The Ki-67 staining was weak and the VEGF staining was negative by immunohistochemistry that indicated the caveolin-1 inhibited the proliferation in human breast cancer MCF7 cell. Conclusion The caveolin-1 might inhibit the growth and proliferation in human breast cancer MCF7 cell through suppressing activation of VEGF signaling pathway.
ObjectiveTo investigate the expressions of contactin-1 (CNTN-1), vascular endothelial growth factor-C (VEGF-C), and its receptor VEGFR-3 (Flt-4) in primary gastric cancer and to explore the relevance among them and their correlation with clinicopathologic features of gastric cancer. MethodsThe VEGF-C, VEGFR-3, and CNTN-1 protein expressions of tumor tissues and normal gastric mucosa tissues in 68 patients with primary gastric cancer were analyzed by immunohistochemistry. The Flt-4-positive vessel density (FVD) and lymphatic vessel density (LVD) were also analyzed by VEGFR-3positive and D2-40-positive staining, respectively. ResultsThe positivity rate of VEGF-C, VEGFR-3, and CNTN-1 protein expression in the primary tumor was 57.4% (39/68), 60.3% (41/68), and 55.9% (38/68), respectively, which was significantly higher than that in the normal gastric mucosa tissues 〔20.6% (14/68), 23.5% (16/68), and 16.2% (11/68)〕， P=0.000. The expressions of VEGF-C, VEGFR-3, and CNTN-1 protein were significantly correlated with TNM stage, lymphatic vessel invasion, and lymph node metastasis (Plt;0.05). The expression of CNTN-1 protein was significantly correlated with VEGF-C (r=0.372, P=0.002) and VEGFR-3 protein expression (r=0.308, P=0.011). In tumor tissues of sixtyeight patients the FVD was (10.41±9.38)/HP, which was significantly lower than LVD 〔(18.19±7.44)/HP〕, P=0.000. Elevated FVD and LVD was significantly found in patients with tumor characterized by later TNM stage, severer lymphatic vessel invasion, and severer lymph node metastasis (Plt;0.05). The FVD of tumor was significantly correlated with VEGF-C (P=0.029) and CNTN-1 protein expression (P=0.003). The LVD of tumor was not significantly correlated with CNTN-1 (P=0.727), VEGF-C (P=0.173), and VEGFR-3 protein expression (P=0.924). The patients with positive expression of VEGF-C, VEGFR-3, and CNTN-1 protein showed poorer prognosis (Plt;0.05). ConclusionsElevated expression of CNTN-1 protein is observed in primary gastric cancer and correlated with VEGF-C and VEGFR-3 protein expression, indicating that combined detection has great value in prediction of invasive potential and prognosis. VEGF-C-mediated CNTN-1 overexpression may promote lymphatic invasion via lymphangiogenesis pathway in patients with gastric cancer.
【Abstract】ObjectiveTo investigate whether tumor necrosis factor-α (TNF-α) enhance the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9(MMP-9) in hepatic cancer cell line HepG2 or not. Methods Cultured HepG2 cells were treated by TNF-α with various concentration and time. The morphological changes of HepG2 cells were studied microscopically and the proliferation of HepG2 were detected by methyl thiazolyl tetrazolium (MTT). The expression of VEGF and MMP-9 mRNA in cultured HepG2 were determined by relative quantitative reverse transcription polymerase chain reaction. The VEGF and MMP-9 protein level in supernatants and in cytoplasm were determined by enzymelinked immunosorbent assay (ELISA) and by immunocytochemical staining, respectively.Results There was a little morphological changes in HepG2 with TNF-α treatment, but no change of cell proliferation in corresponding time. The expression of VEGF and MMP-9 mRNA was enhanced gradually with the TNF-α concentration increasing, the VEGF and MMP-9 protein level in supernatants and in cytoplasm was elevated gradually with the concentration increasing. There was a dependance on the concentration when the concentration of TNF-α was lower than or equal to 104 U/L. Furthermore, the effect of promotion was close to peak when the TNF-α concentration up to 104 U/L; but no timeeffect pattern observed. Conclusion TNF-αJP can enhance the expression of VEGF and MMP-9 at the level of mRNA and protein in hepatic cancer cell line.
Objective To investigate the effects of simvastatin on pulmonary function and vascular endothelial growth factor ( VEGF) levels in induced sputumof patients with COPD exacerbation( AECOPD) .Methods Thirty-eight patients with AECOPD were divided into two groups randomly, ie. a routine medical treatment( RT) group( n =30) and a routine + statin medical treatment( RST) group( n =28) . The VEGF levels in serumand induced sputum were detected by ELISA on the first day and after a week treatment in hospital, respectively. Meanwhile, the pulmonary function measurements were performed. Results There were no significant differences in the pulmonary function ( FEV1% pred and FEV1 /FVC) and VEGF levels in induced sputumbetween the two groups before treatment( P gt;0. 05) . The RT group showed no significantchanges in any parameters before and after a week treatment( P gt; 0. 05) . FEV1% pread, FEV1 /FVC and VEGF levels in induced sputum in the RST group after a week treatment significantly increased compared with those before treatment and the RT group( P lt;0. 01, P lt;0. 01, P lt;0. 05) . But There were no significant differences in serumVEGF levels between the two groups before and after a week treatment. The VEGF levels in induced sputum were positively correlated to FEV1% pread and FEV1 /FVC after a week treatment( r =0. 430, P lt;0. 05; r = 0. 388, P lt; 0. 05) . Conclusions Simvastatin may reduce the decline in pulmonary function and decrease the levels of VEGF in induced sputum of patients with AECOPD. Improvement in pulmonary function may be related to down-expression of lung VEGF
Objective To investigate whether ADAM33 ( A disintegrin and metalloproteinase 33) gene polymorphismhas effect on the airway inflammation of COPD. Methods A total of 312 COPD patients were recruited for this study. Four polymorphic loci ( T2, T1, S2, and Q-1) of ADAM33 were selected for genotyping by using the polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP) method. Total and differential cell counts, contents of TNF-α, IL-6, IL-8, and VEGF in induced sputumwere detected. The relationship between genotypes and inflammatory reaction was analyzed. Results On locus T2, the cell counts and content of TNF-αin induced sputum increased significantly in the carriers with GG genotype than those with AA and AG genotypes ( Plt;0.01 and Plt;0.05) . On locus T1, the lymphocyte counts in induced sputumincreased significantly in the carriers with GG genotype than those with AA and AG genotypes ( Plt;0.05) ; but the content of IL-8 in induced sputumwas higher in AA and AG genotypes ( Plt;0.05) . On locus Q-1, the contents of VEGF and IL-8 in induced sputum increased significantly in the carriers with GG genotype than those with AA and AG genotypes (Plt;0.05) . On locus S2, the total cell counts in induced sputumincreased significantly in the carriers with GG genotype than those with CC and CG genotypes ( Plt;0.05) , and the content of IL-8 in induced sputum increased significantly in GG genotype ( Plt;0.01 ) . Conclusion These results suggest that ADAM33 polymorphism may participate the pathogenesis of COPD by promoting airway inflammation.
Objective To observe the effect of vascular endothelial growth factor (VEGF) in fracture healing and to investigate the influence of VEGF and VEGF antibody in fracture healing. Methods One hundred and five rabbits were used tomake fracture model in the left radius and randomly divided into control group,VEGF group and VEGF antibody group. VEGF and VEGF antibody were used in the VEGF group and VEGF antibody group respectively, then the blood flow of the fracture ends was measured by single photon emission computed tomography (SPECT) 8,24 , 72 hours, 1, 3, 5 and 8 weeks after fracture, the X-ray films of the fracture sites were taken after 1, 3, 5 and 8 weeks to observe the fracture healing. Results The blood flow of the fracture ends in the VEGF groupincreased during aperiod from 8h to 3wk after fraction when compared with that of the control group, and no obvious difference was seen on the X-ray films between the two groups. In the VEGF antibody group, the blood flow of the fracture ends decreased obviously when compared with that of the control group. The fracture healing processwas interfered seriously and nonunion change was seen in the fracture site. Conclusion The lack of VEGF will interfere with the fracture healing process and result in nonunion in the fracture site. Administration of ectogenous VEGF may promote fracture healing by increasing the blood flow of the fracture ends.
Objective To evaluate an effect of the vascularendothelial growth factor (VEGF) geneactivated matrix (GAM) on repair of the sciatic nerve defect in rats. Methods The peripheral nerve extracellular matrix（ECM） was harvested by the chemical extraction from 30 SD rats. The VEGF-GAM comprised of ECM and the plasmids encoding VEGF. Thirty adult Wistar rats were made as a model of the asciatic nerve defect and were randomly divided into the following 3 groups(n=10): Group A (VEGF-GAM conduits), Group B (ECM conduits),and Group C (autografts). At 12 weeks, the rats from each groupwere subjected to an inspection for the walking tract analysis and electrophysiological and histomorphological studies.Results The VEGF DNA could be retained in GAM, promoting the transgene expressing in the sciatic nerve, and more importantly, in the axotomized neurons in the spinal cord for 12 weeks. The motor neuron recovery rate in Group A (79.13％±2.53％) was similar to that in Group C (75.26％±4.48％, Pgt;0.05), but significantly better than that in Group B (56.09％±1.89％, Plt;0.01). The number of the regenerationaxons in the distal sciatic nerve in Group A (13 463±794/mm2) was significantly lower than that in Group C (16 809±680/mm2, Plt; 0.01), but significantly higher than that in Group B (10 260±1 117/mm2,Plt;0.01). The motor nerve conduction velocity in Group A (16.44±1.65 m/s) was significantly lowerthan that in Group C (23.79±2.75 m/s, Plt;0.01), but significantly higherthan that in Group B (12.8 ±1.42 m/s, Plt;0.01). The recovery rate of thegastrocnemius muscle wet weight in Group A (71.40％±3.05%) was significantlylower than that in Group C (87.00％±1.87％,Plt;0.01), but significantly higher than that in Group B (50.00％±4.90％, Plt;0.01). The sciatic nerve function index in Group A （39.37%±4.81%） was significantly lower 〖KG6〗than that in Group C (26.27%±2.71%, Plt;0.01), but significantly higher than that in Group B (4693％±296％, Plt;0.01). Conclusion The results indicate that VEGF-GAM as a bridge can promote the functional recovery of the defected sciatic nerve in rats, but the effect is not so good as that by autografts.