Protective Effect Of Salvianolate On Human Umbilical Vascular Endothelial Cells Induced By High Glucose

Objectives:To investigate the protective effect of Salvianolate on the injury of human umbilical vein endothelial cells induced by high glucose in vitro, and to explore its underlying mechanism.Methods:1. The model of HUVEC induced by high glucose in vitro Human umbilical vein endothelial cells was cultured in different dulbeeeo's modified egle medium(DMEM) different times(0hour,24hours,48hours,72hours). The experiment was randomly divided into Control group(concentration of glucose: 5.5mmol/L) and 6 high glucose groups which were injured by glucose with different concentration (10mmol/L, 20mmol/L, 30mmol/L ,40mmol/L ,50mmol/L ,respective- ly) .After different time, the cell vitality,the activity of GSH-PX,MDA content in supernatant were measured, and the morphological changes of HUVEC were observed by inverted phase contrast microscope.2.Protective Effect of Salvianolate on Human Umbilical Vascular Endothelial Cells Induced by High GlucoseThe experiment was divided into 5 groups: Control group(concentration of glucose:5.5mmol/L), high glucose group(concentration of glucose:30mmol/L), low dose Salvianolate group(30mmol/L glucose + 50mg/L Salvianolate), medium dose Salvianolate(30mmol/L glucose + 100mg/L Salvianolate),high dose Salvianolate (30mmol/L glucose + 200mg/L Salvianolate).The HUVEC was cultured in different cell culture fluid for 48hours, the cell vitality,the activity of GSH-PX,MDA content,the secretary content of nitric oxide(NO)and endothelin-1(ET-1)in supernatant were measured. Result:1. The model of HUVEC induced by high glucose in vitro:1) The cell vitality(OD optimum) was gradually increased as cultured time goes by in 72 huors,but at the same cultured time,compairing with control group(5.5mmol /L), in 10mmol/L,20mmol/L groups ,the increasing of cell vitality(OD optimum) was inhibited obviously (P=0.003);However, the cell vitality(OD optimum) was gradually decreasing as cultured time goes by in 72 huors. (P=0.02);2) As cultured time goes by ,the MDA content was increasing, but at the same cultured time (excepting 0 hour), compairing with control group(5.5mmol /L), the MDA content was increased obviously by the concentration of glucose advancing(P=0.01);3) As cultured time goes by , the activity of GSH-PX was increasing, but at the same cultured time (excepting 0 hour), compairing with control group(5.5mmol /L), the activity of GSH-PX was decreased obviously by the concentration of glucose advancing(P=0.02);4) Compairing with control group(5.5mmol /L), from inverted phase contrast microscope, the cell shape of 10mmol/L,20mmol/L groups cultured 48 hours was without significant distinction. However,the cell shape of 30mmol/L,40mmol/L,50mmol/L groups were significantly different from the control group, the cell shape of 30mmol/L,40mmol/L groups distorted while the cell boundary was still clear,and compairing with the 30mmol/L group,40mmol/L group was distorted obviously,the cell shape of 50mmol/L groups was distorted apparently and the boundary was not clear, and with dead cells fall off from the bottom of wells.2. Protective Effect of Salvianolate on Human Umbilical Vascular Endothelial Cells Induced by High GlucoseCompairing with control group(5.5mmol /L),the cell vitality(OD optimum),the GSH-PX content and secrete of nitric oxide(NO) was decreased obviously (P=0.003,P=0.02,P=0.01).However, the MDA content and the secrete of Endothelin-1 was increased obviously (P=0.01,P=0.003);Meanwhile, in Salvianolate group ,compairing with high glucose group(30mmol /L), the cell vitality(OD optimum),the GSH-PX content and the secrete of nitric oxide(NO) was advancing obviously (P=0.02,P=0.01,P=0.003),the MDA content and the secrete of Endothelin-1 was descending obviously (P=0.03,P=0.004).Conclusion:1.High glucose can induced the cell viability descending, active oxygen produced increasing and the activity of antioxidase descending,and caused HUVEC injury, and the extent of cellular damage depends on the concentration of glucose and the cultured time.The condition of 30mmol /L and cultured 48 hours is suitable to study the protective effect of Salvianolate on HUVEC induced by high glucose.2. The Salvianolate can protect the injury of HUVEC induced by high glucose in vitro,and the mechanism may be associated with scavenging reactive oxygen radicals,reducing lipid peroxidation,and increasing the activities of intracellular antioxidase,inhibiting the secrete of Endothelin-1.