Experimental Study On Inhibition Of Proliferation Of Human Umbilical Vein Endothelial Cells By Spica Prunellae And Spina Gleditsiae

ObjectiveDiabetic retinopathy(DR) is the capillary pathology concerning to the diabetic. Developed to a certain stage,there will be neovascularization,it damage visual function Seriously,the reason is not exactly known,therefore there is no ideal drug treatment.Angiogenesis dependent on the interaction of vascular endothelial cells and extracellular matrix,So at present vascular endothelial cells have been the unique target cells on treatment of anti-angiogenesis.In this study,we investigate the proliferative inhibition of vascular endothelial cells by Spica Prunellae and Spina Gleditsiae and further study their mechanism of the proliferative inhibition of vascular endothelial cells from PCNA(Proliferating cell nuclear antigen),in order to provide scientific basis for pursuing safe and effective natural drugs to prevent and cure neovascularization in DR.MethodsHuman umbilical vein endothelial cells(HUVEC) were cultured and were used for experiment in good condition.HUVEC proliferation were induced by 100umol/L CoCl₂,in the same time 10mg/ml,20mg/ml,40mg/ml Spica Prunellae and 20mg/ml,40mg/ml,80mg/ml Spina Gleditsiae were added to the HUVEC separately.The following studies were made after 24 hours incubation.1.The effects of proliferative inhibiton by Spica Prunellae and Spina Gleditsiae on the HUVEC were assayed with MTT and CCK-8 methods.2.The changes of proliferating cell nuclear antigen(PCNA) expression of HUVEC regulated by Spica Prunellae and Spina Gleditsiae were assayed with flow cytometry(FCM). Results1.100umol/L CoCl₂ can induce the proliferation of HUVEC.2.COCl₂ can upregulate the expression of PCNA in HUVEC.3.Spica Prunellae and Spina Gleditsiae can significantly inhibit the proliferation of HUVEC induced by CoCl₂ and these effects were dose dependent.4.Spica Prunellae and Spina Gleditsiae can downregulate the expression of PCNA in HUVEC and this decrease was dose dependent.Conclusion1.The proliferation of HUVEC induced by CoCl₂ may be contributed to upregulating PCNA expression.2.Both Spica Prunellae and Spina Gleditsiae can markly inhibit the HUVEC proliferation induced by CoCl₂.3.The mechanism of inhibiting proliferation by Spica Prunellae and Spina Gleditsiae in HUVEC may be contributed to downregulating PCNA expression and hindering the DNA synthesis of HUVEC.4.The inhibitive effects of Spica Prunellae and Spina Gleditsiae on the proliferation make them probably become reliable and effective drugs to prevent and cure neovascularization in DR.