| Objective:To observe the effect and relationship of gobular adiponectin on the highglucose-induced NO Level and DDAH-ADMA system in HUVECs.Methods:The HUVECs was treated to the logarithmic growth phase in vitro, andgad(2.5μg/mL) was administered in the high glucose-induced of them for differenttime(24,48hours),and then the HUVECs was treated with gad of0.1μg/mL,0.5μg/mL,2.5μg/mL,12.5μg/mL in high glucose environment for48respectively.The nitratereductase method was to detect the NO concentration of culture supernatant;HUVECs were randomized to four groups:normal control(5.5mmol/L glucose),highglucose(33.0mmol/L),gad(33.0mmol/L glucose+2.5μg/mL gad) and L-NAME (33.0mmol/L glucose+2.5μg/mL gad+100μmol/L L-NAME).The nitrate reductase methodwas to detect the NO concentration of culture supernatant and the chemical colorimetricanalysis was to analyse the activation of NOS and then the level of ADMA was detected by HighPerformance Liquid Chromatography in experimental group and normal control group,The activation of DDAH was reflected by the level of ADMA, and both eNOS andp-eNOS were analysed by Western blot in experimental group and normal controlgroup.Results:①HVUECs was induced by high glucose,and two time points(24hours later and48hours later) were set in order to observe the concentration of NO in HVUECs.Theconcentration of NO was increased after24hours, but decreased after48hours in thehigh glucose group. Compared with the high glucose group, the concentration of NOincreased extremely in the gad group （P<0.05）②Different concentration of gad(0.1,0.5,2.5,12.5μg/mL) was given to the high glucose-induced(48hours)HUVECs,the level of NO was first increased in correspondence with the elevatedconcentrations of gad, but there was no difference between groups of theconcentrations of2.5μg/mL and12.5μg/mL of gad;③2.5μg/mL gad was given in the48hours high glucose-induced HVUECs, compared with the normal control group,the concentration of NO, as well as the activation of NOS and the expression ofp-eNOS(P<0.05), decreased sharply in the high glucose group. but there was nodifference in the expression of the total eNOS among them(P>0.05).④Compared withnormal control group,high glucose induced the increasing of ADMA, and the decreasingof the activation of DDAH. The differentia had remarkable significance(P<0.05).Compared with the high glucose group, the concentration of NO, as well as theactivation of NOS and the expression of p-eNOS(P<0.05),greatly increased in the gadgroup.and the expression of eNOS was the same among them (P>0.05);comparedwith the high glucose group, the level of ADMA was decreased, and the activation ofDDAH was increased in the gad group. The differentia had remarkable significance(P<0.05).Compared with the gad group, the concentration of NO, as well as theactivation of NOS and the expression of p-eNOS,was decreased in the L-NAMEgroup, The differentia had remarkable significance(P<0.05), and the expression ofeNOS was the same between them (P>0.05).,What’s more, the level of ADMAincreased and the activation of DDAH decreased.The differentia had remarkablesignificance(P<0.05).Conclusions:1. In high glucose environment gad enables to reverse the decreasing of NOwhich is synthesized by HVUECs, and to increase the activation of NOS andphosphorylation of eNOS-Ser1177,but there is no difference in the expression ofeNOS.2. In high glucose environment gad enables to increase the activation of DDAH,and decrease the level of ADMA.The reason why this effect is brought about may bethat gad induces the increasing of NO and restores the activity of NOS.Simultaneously,the increasing activation of DDAH and decreasing level of ADMA could increase the level of NO and the activation of NOS. |
