Experimental Study On The Inhibition Effect And The Mechanism Of Mimeng Flower Decoction On The Proliferation Of Human Umbilical Vein Endothelial Cells Induced By Chronic Hypoxia In Vitro

ObjectiveTo study the inhibition effect of Mimeng Flower Decoction on the proliferation of human umbilical vein endothelial cells (HUVEC) induced by chronic hypoxia in vitro, and further study the mechanism of the inhibition effect by using immunocytochemistry and western blot methods to test the expression of hypoxia inducing factor (HIF-1α), vascular cell adhesion molecule (VCAM-1),fibronectin (FN) in HUVEC regulated by Mimeng Flower Decoction; in order to conclude the inhibition effect of Mimeng Flower Decoction on neovascularization and provide experimental basis for Mimeng Flower Decoction used in clinic to prevent and cure diabetic retinophty(DR).MethodsHuman umbilical vein endothelial cells (HUVEC) cultured in vitro which were in good condition, were induced by 100umol/L CoCl2, in the same time 10mg/ml,20mg/ml,40mg/ml Mimeng Flower Decoction extra were added to the HUVEC separately.The following studies were made after 24 hours incubation.1. Through MTT method to check up the inhibition effect of Mimeng Flower Decoction on the proliferation of human umbilical vein endothelial cells (HUVEC) induced by chronic hypoxia in vitro.2. Through immunocytochemistry and werstern blot methods to test the expression of hypoxia inducing factor (HIF-1α), vascular cell adhesion molecule-1(VCAM-1),fibronectin(FN)regulated by Mimeng Flower Decoction in HUVEC induced by chronic hypoxia in vitro.ResultsCells grew more rapidly in hypoxia group than in Mimeng Flower Decoction groups.1.The results of MTT test show that the hypoxia group can increase the optical density(OD) values compared with blank group(P<0.01);Mimeng Flower Decoction groups can reduce the OD values obviously compared with hypoxia group(low dose group:P<0.05; high and medium dose group: P<0.01);and these effects were dose dependent (P<0.01); the high dose of Mimeng Flower Decoction group'OD values were the lowest.2. The results of immunocytochemistry test show that the expression of HIF-1α, VCAM-1, FN increased in hypoxia group compared with blank group (P<0.01); Mimeng Flower Decoction could down regulate the expression of HIF-1α, VCAM-1, FN compared with hypoxia group;and the expression of HIF-1α, VCAM-1, FN reduced in high dose group compared with medium and low dose group。3. The results of werstern blot test show that the expression of HIF-1α, VCAM-1, FN increased in hypoxia group compared with blank group. Mimeng Flower decoction could down regulate the expression of HIF-1α, VCAM-1, FN compared with hypoxia group.Conclusions1.CoCl2 can induce proliferation of HUVEC.2. Mimeng Flower Decoction can markly inhibit the proliferation of HUVECin hypoxia state, and the effects of inhibition were dose dependent.3. Mimeng Flower Decoction can inhibite the proliferation of HUVEC, reduce the injury of cell and improve the function of the cell possiblely by reducing the expression of HIF-1α, VCAM-1, FN in HUVEC in hypoxia state.