Vascular endothelial cell(VEC) is a kind of simple squamous epithelium lined on the inner surface of blood vessels. VEC is an important barrier between the blood and tissue and it also plays a key role in regulating inflammation, thrombosis, endothelial cells mediated vasodilatation and endothelial regeneration. These processes should be controlled by a variety of complex mechanism which requires us to find out. With results of the researches in vascular endothelial cell function, the important roles that microRNA in vascular endothelial cell function draws more and more researchers' attention. MicroRNAs control gene expression in post-transcriptional level and affect the function of endothelial cells. This review focuses on the research progress on regulatory mechanism of microRNA to endothelial cell inflammation, thrombosis, vasodilation and endothelium regeneration.
ObjectiveTo observe the effects of A549 cells under hypoxicconditions on the migration of human umbilical vein endothelial cells (HUVECs) and microvascular formation. MethodsAfter cultured for 24 h in normoxia condition(21% O2),hypoxia condition (2% O2),and anaerobic condition (0% O2),respectively,morphology of A549 cells was observed with inverted phase contrast microscope,proliferation was detected by MTT assay,and intracellular hypoxia-inducible factor-1α (HIF-1α) protein was detected by immunocyto-chemical technique,for determining whether the hypoxia model is successful. Then A549 cells' supernatant in the normoxic group,the hypoxia group and HUVECs culture medium were taken to intervene HUVECs. The migration of HUVECs was observed with cell scratch test,pseudopodia formation of HUVECs was observed with microfilament green fluorescent staining method,and blood vessel formation was observed with three-dimensional culture techniques in vitro. ResultsCompared with the normoxic group,the growth of A549 cells was better in the hypoxia group with more proliferation,and was poor in the anaerobic group with decreased number of cells. A549 cells in the hypoxia group and the anaerobic group both expressed HIF-1α protein,which was more obvious in the anaerobic group. Compared with the HUVECs supernatant intervention group,the hypoxia supernatant intervention group and the normoxic supernatant intervention group both had varying degrees of migration,pseudopodia structure formation and vascular lumen sample structure formation,which were more obvious in the former group. ConclusionA549 cells in hypoxic environment grow very well,proliferated significantly,but anaerobic environment is not conducive to the growth of A549 cells which found to be apoptosis. A549 cells in hypoxic environment can promote HUVECs migration,pseudopodia formation and angiogenesis.
Objective To study the feasibility of transplanting autologous venous endothelial cells, as the liner, to the allogenic vein and to investigate the patency rate after such transplantation. Methods Autologous endothelial cells were gained after the administration of 0.2% collagenase and the centrifugalization of the enzyme liquid. The cells were not cultivated in a 60 ml plastic culture until the presence of the second generation. The cultivated cells were confirmed as endothelial cells by factor Ⅷ related antigen. The multiplied cells were lined in vitro onto the luminal surface of allogenic vein that was disposed by freeze-drying and radiation. The orthotopic transplantation of autologous venous endothelial cells was performed after the 9-day incubation. Results (9.47±0.35)×106 endothelial cells were obtained after the cultivation. Three hours after cell seeding, the luminal surface of allogenic vein was covered with vast endothelial cells but still had not formed an intact endomembrane. On day 9, the luminal surface was covered with a continuous endothelial monolayer and the arrangement and the shape of the cells all showed the perfect condition of endothelial cells. Eight weeks later, all the transplanted veins kept unobstructed. Conclusion The approach of lining allogenic vein with autologous endothelial cells in vitro may keep the vein unobstructed in the long term.
bjective To investigate the correlation between expression of vascular endothelial growth factor(VEGF) of serum and tumor tissues and the clinical prognosis in patients with breast cancer. Methods The expressions of VEGF level of serum and tumor tissues in 44 patients with invasive duct breast cancer, 13 with benign breast diseases and 40 healthy controls. Serum VEGF level was measured by ELISA method. The protein expression of tissue VEGF, ER and C-erbB-2 were evaluated by immunohistochemistry LSAB method. Results Serum VEGF level and tissue VEGF expression in breast cancer were higher than those in benign breast diseases （P＜0.001）, and there was no significance in benign breast diseases and healthy controls （Pgt;0.05）. VEGF expression was correlated with lymph node metastasis (P＜0.01), ER and C-erbB-2 expression (P＜0.05, P＜0.01) and clinical stage （P＜0.01）. There were no statistical correlation between VEGF expression and age, tumor size (Pgt;0.05). Conclusion There is positively correlation between serum VEGF level and tissue VEGF expression, and between VEGF expression and clinic prognosis. Serum VEGF level may be one of important index of prognosis estimation in patients with breast cancer.
Objective To discuss the endothelial cell which was modified by exogenous anticoagulant genes contribute to the increase of antithrombosis activity of lined vascular prosthesis and the influence to other physiological functions of endothelial cells. Methods This summarized paper was made on literature review of recent years. Results The transfection of genes, including plasminogen activator (tPA, uPA, Urokinase), thrombomoduline (TM) and hirudin, etc, to endothelial cells resulted in not only the increase of antithrombosis activity of local vascular, but also the decrease of endothelial cell function in adherence and proliferation. Conclusion The increase of antithrombosis activity of lined vascular prosthesis has been done by exogenous genes. However, this technique ought to be studied, intensively.
ObjectiveTo summarize the progress on the injury mechanism of vascular endothelial cells in atherosclerosis.MethodsThe latest progress was reviewed in recent literatures.ResultsAll kinds of etiological factors have activated NF-kappa B and cytokines in the development of atherosclerosis, which lead to expression of cell adhesive molecules and adhesion of monocytes to vascular endothelial cells.A variety of inflammatory mediums are released, which can directly damage endothelial cells.Besides, the inflammatory mediums make monocytes and neutrophils attach to endothelial cells by immune mechanisms, which injure the endothelial cells more severely. Meanwhile the damaged membrance structure leads to the production of AECA which activates the complementary system. Then the vascular endothelial cell injury is aggravated and the development of atherosclerosis accelerated. ConclusionIt is very important to recognize the injury mechanism of vascular endothelial cells in the development of atherosclerosis for prevention and treatment of atherosclerosis.
ObjectiveTo study the early functional change of sinusoid endothelial cell after liver transplantation in rat, and to investigate the endothelia protective effect of prostaglandin E1(PGE1). MethodsRat orthotopic liver transplantation model was performed in “twocuff method”, grouped as follows: group A served as normal rat blank control, group B as operative control with normal donor, group C as experimental control with shock donor, and group D as experimental group with shock donor and PGE1 administration (n=8 in each group). Transplanted groups (referring to recipients without specific definition) were sacrificed 6 h after operation for blood taken to detect serum liver enzymes (ALT, LDH), malondialdehyde (MDA), nitric oxide (NO) and plasm endothelin (ET). Liver tissue was resected at the same time for standard pathologic examination. Comparison of the difference the results was made between groups. ResultsCold preservation time and anhepatic phase were similar in each group, (2±0.5) h and (15±3) min respectively. All survived 6 h after transplantation (8/8) in group B and D with a survival rate of 100%, only 5 survived 6 h after transplantation in group C (5/8) with a survival rate of 62.5%. Comparing with group C, blood ALT, LDH, MDA, ET decreased and NO increased significantly in group D (Plt;0.05). Marked histologic structural damage was observed in group C, while normal light microscope appearance was better preserved in group C and D. ConclusionMarked sinusoid endothelia injury occurs during liver transplantation. Concentration of serum NO and plasm ET well presents its function. PGE1 relieves endothelia injury by improving hemodynamics and stabilizing sinusoid endothelial cell plasma membrane.
ObjectiveTo investigate the expression of vascular endothelial growth factor （VEGF） and proliferating cell nuclear antigen (PCNA) in colorectal cancer and its relationship with metastasis and recurrence. MethodsParaffinembedded specimens from 59 patients with colorectal cancer, 16 patients with adenomas and 12 normal colonic tissues were examined and compared by SP immunohistochemical method. ResultsThe positive rate of VEGF in colorectal cancer were significantly higher than that in adenomas (P＜0.05). The positive rate of VEGF in Dukes A and B stage of colorectal cancer were significantly higher than those in Dukes C and D (P＜0.05). Expression of VEGF in postoperative recurrence group was markedly higher than that in the group with no recurrence (P＜0.05). Proliferative activity expression suggested that the poorer the differentiation, the more PCNA increased in case of lymphnode or hepatic metastasis. The PCNA showed marked difference between postoperative and nonpostoperative recurrences (P＜0.05). Conclusion The expression of VEGF and PCNA is closely related to the invasion and metastasis of tumor during the operation. The increased VEGF and high PCNA implies that there may be some potential metastasis present.
ObjectiveTo explore the relation between vascular endothelial growth factor （VEGF） and the formation of tumor thrombosis in the main trunks of portal vein (PVTT). MethodsTumor specimens were collected from 36 patients (16 patients with PVTT, the other patients without PVTT and metastasis) undergoing resection of hepatocellular carcinoma (HCC) and portal thrombectemy, PVTT specimens of 16 patients named group A1, the same patients’ with HCC named group A2, tumor specimens of the other patients named group B. In situ hybridization and immunohistochemistry were used to investigate VEGF mRNA, protein and microvessel density (MVD) on surgical specimens. The intensity was evaluated using a computer image analyzercell analysis system.ResultsVEGF mRNA expression was detected in the tumor’ cell of the specimens. The expression rates of VEGF mRNA in the group B, A2, A1 were 30%, 100%, 100% respectively, and the expression rates of VEGF mRNA in group A2 and A1 were higher than that in group B （P＜0.01）. The intensity of VEGF mRNA in group A2 （0.078 5±0.019 6） were lower than in group A1 （0.194 4±0.059 0） （P＜0.01）. VEGF protein expression was often detected in the tumor cell, vascular endothelial cell and fibroblast cells. Invasion was detected in small vein in group A2, more tumor cell colony detected in group A1. The expression rates of VEGF protein in group B, A2, A1 were same as VEGF mRNA; the intensity of VEGF protein in A1 （0.165 6± 0.034 5） was higher than in group A2 （0.108 1±0.024 3） （P＜0.01）. MVD in group B, A2, A1 was 31.9±14.4, 63.3±15.1, 116±27.6/view of 200 microscopefield, MVD in group A1 was higher than group A2 （P＜0.01）, higher in group A2 than in group B. There was a statistically significant correlation between the intensity of VEGF expression and MVD in group B,A2 and A1. ConclusionVEGF could play an important role in the invasion, metastasis of HCC and the formation of PVTT. Angiogenesis in tumor is correlated well with the progression of HCC.
Objective To be expressed human vascular endothelial growth factor (VEGF) recombinant protein in Escherichia Coli in high level. Methods VEGF was amplified from human fetal brain cDNA library, the amplified fragment was inserted into M13mP18 and confirmed to be VEGF165cDNA by restriction mapping and DNA sequencing, then it was combined with an expression vector PRL621. This recombinant plasmid overexpressed a 20kd recombinant protein in E.Coli(TG1), the protein was isolated and purifed from E.Coli, and initially renatured. Results The overexpressed recombinant protein was 35% of the total cell protein, the sequence of its first 15-N terminal amino acid was identrical to that of the human natural VEGF protein, Chorioallantoic membrane(CAM) assay showed that the rhVEGF promated new capillary vessels formation. Conclusion The genetic engineering Escherichia Coli can express human vascular endothelial growth factor in high level.