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Objective To evaluate the influence Qizhu decoction on VEGF and its receptor KDR/flk-1 protein expression in MGC803 cell. Methods Four groups of MGC-803 Cells were established, and intervened with blank control, DMSO control, high dose Qizhu decoction, and low dose Qizhu decoction respectively. VEGF and KDR/flk-1 protein were detected by flow cytometry and western bloting. Results The value of VEGF expression was 120.0±10.8, 116.8±14.7, 95.0±12.5, and 108.4±13.5 respectively in each group. The difference between high dosage Qizhu decoction group and the bland control group was significant. Value of KDR/flk-1 were 10.4±3.5, 9.0±3.4, 6.8±2.3, and 6.8±3.5 in each group respectively. There was no significant difference among these 4 groups. The grayseale values of VEGF expression was 14.45±4.61, 12.32±3.27, 2.58±0.84, and 3.45±1.12 in each group respectively. There was significant difference between the QiZhu groups and the blank control group. Meanwhile grayseale values of KDR/flk-1 expression were 3.87±1.05, 3.55±1.32, 3.62±1.01, and 3.73±0.88 in each group respectively, showing no significant difference among 4 groups. Conclusion Rough extraction of QiZhu decoction down-regulated the protein expression of VEGF, but had no effects on the expression of KDR/flk-1.
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· AIM :To investigate the role of pericytes in growth of retinal microvascular endothelial cells with a co-culture system in order to understand some mechanism of angiogenesis in hypoxia induced retinal neovascular disorders.(RMECs) were isolated by a modified protocol using CD31 coated Dynabeads, and identified by immunocytochemical staining with anti-Factor Ⅷ and CD31 antibodies. Rat retinal pericytes were isolated and characterized by immunofluorescent staining with PDGFR-β; and desmin antibodies. Pericytes and RMECs were cultured in a contact co-culture system both under normoxia and hypoxia by Millicell chamber. RMECs proliferation was evaluated by MTT and cell cycle assay with flow cytometry. RT-PCR was used to detect the alteration of KDR/Flk-1 mRNA level in RMECs under normoxia or hypoxia in the co-culture system.harvested with the modified isolating method. The two cell types were identified by positive Factor Ⅷ, CD31 and PDGFR-β, desmin cytochemical staining respectively.RMECs proliferated significantly under hypoxia from 3 to 9d with a maximal rate on day 6 (24.9%, P ＜ 0.01) by MTT. In the co-culture system, the proliferation of RMECs was inhibited by pericytes. After 6d exposure to hypoxia,the fraction of S-phase RMECs number was greatly increased by 43.9% (P ＜ 0.01). In the co-culture system,RMECs proliferation was inhibited by pericytes through decreasing the fraction of S-phase cell number both under normoxia (3.6%, P＜0.05) and under hypoxia (15.1%,P＜0.01). KDR/Flk-1 mRNA level in single cultured RMECs was shown to increase approximately 1.3-fold when exposed to hypoxia. Compared with single cultured RMECs, co-culture with pericytes could decrease KDR/Flk-1 mRNA by 45.1% (P＜0.05) and 27.7% (P ＜ 0.05) under normoxia and hypoxia condition respectively.pericytes could inhibit proliferation of RMECs under both normoxia and hypoxia. The inhibition effects of pericytes maybe, at least in part, due to downregulation of KDR/Flk-1 of RMECs. These findings confirm that pericytes could be a potential inhibitor in the pathogenesis of retinal neovascularization.
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Objective To study the anti-agiogenesis effect of Taohong Siwu Decoction Ⅱ(THSDⅡ)and to explore its mechanism.Methods Chicken Chorioallantoic Membrane(CAM)assay was used to study the anti-agiogenesis action of THSDⅡin vivo.MTT assay was used to investigate its effect on the proliferation of human umbilical vein vascular endothelial cells ECV304 in vitro.Immunohistochemistry assay was used to observe its effects on the expression of KDR/FLK-1 in endothelial cells and the amount of micro-vessel density(MVD)in mice with B16 melanoma.Results THSDⅡ at the dosages of 1 g / mL and 2 g / mL could obviously inhibit agiogenesis in the CAM(P
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AIM: We observed the expression of KDR/Flk1 in post-myocardiac ischemia(MI) SD rats in order to explain the effect of captopril and its relationship with myocardium angiogenesis after long-term administration.METHODS: The MI model was made by LAD ligation.Captopril was administered for 6 weeks.Immuohistological method and FQ-PCR were used to test the myocardium KDR/Flk-1 expression.RESULTS: In captopril group,no inhibitory effect was observed in myocardium factor VIII expression,but KDR/Flk-1 decreased.The copies of KDR/Flk-1 mRNA reduced dramatically when compared to control group,false-operation and normal group(P0.05).CONCLUSION: ACEI down-regulates KDR/Flk-1 and its mRNA expression in ischemic rat myocardium after long-term administration of captopril,but does not inhibit angiogenesis.So we suspect that some other pathways exist,which can not affect by ACEI,or that ACEI just reduces abnormal angiogenesis.