The Effects Of Recombinant Adiponectin In Endothelial Cells Induced By Oxidative Stress Via NF-κ B-iNOS-nitric Oxide Pathway

OBJECTIVE:To investigate the effects of NF-κB,iNOS and NO in human umbilical vein endothelial cells induced by Tert-butyl hydroperoxide.Further,to explore the effects of recombinant adiponectin in human umbilical vein endothelial cells induced by oxidative stress via NF-κB-iNOS-nitric oxide pathway.METHODS::Human umbilical vein endothelial cells were cultured in vitro.cells viability was measured by MTT assay.Nitric oxide levels were measured by Griess assay.Inducible nitric oxide synthase(iNOS) protein and NF-κBp65 fragment were detected by Western blot.After treatment with L-NAME 600umol/L for 24 hours,nitric oxide levels were measured by Griess assay.Further,cells were pretreated with recombinant adenovirus containing adiponectin(MOI100) for 48 hours,investigate the effects of recombinant adiponectin in human umbilical vein endothelial cells induced by t-BHP.RESULTS:Cells viability was reduced in a time-dependent and a dose-dependent manner after exposure of HUVEC to t-BHP at range of 12.5-100umol/L for 30-120min.The levels of cytoplasmic iNOS protein were elevated and the peaking was at 60min.Meanwhile,treatment with t-BHP resulted in nucleus NF-κBP65 fragmentpeaking at 30min.treatment with L-NAME 600umol/L attenuated the levels of Nitric oxide due to t-BHP alone. Further,cells were pretreated with recombinant adenovirus containing adiponectin for 48h,the study showed that recombinant adiponectin decreased the levels of nitrite by t-BHP in HUVEC (p＜0.05).Furthermore,recombinant adiponectin reduced the expression of iNOS protein and NF-κBp65 fragment(p＜0.05).CONCLUSION:cells viability was reduced by t-BHP and the levels of nitrite was increased.The cell signalling of NF-κB-iNOS-nitric oxide mediated tBHP-induced damage in HUVEC.Recombinant adiponectin protected HUVEC from tBHP-induced damage via down-regulation of NF-κB-iNOS-nitric oxide pathway.