Transcriptional Up-regulation Of Adiponectin Receptor 1 Expression In Vascular Smooth Muscle Cells In Rat Mesenteric Artery By Organ Culture

Objective:Cardiovascular disease(CVD)is a group of circulatory system diseases characterized by cardiac or vascular abnormalities.Atherosclerosis(AS)is a chronic inflammatory disease of the vascular wall,and AS is the main pathological basis of CVD.Inflammation is a key link in vascular endothelial injury and vascular smooth muscle cells(VSMCs)dysfunction that leads to AS.Adiponectin signaling pathway plays a role on insulin sensitization,anti-inflammation,anti-atherosclerosis and promoting angiogenesis.Although adiponectin receptor 1(AdipoR1)is the main receptor of adiponectin(APN)in VSMCs,the expression of AdipoR1 in VSMCs and its effect on vascular function under pathological conditions have not been reported.With the help of the organ culture model,the main purpose of this study is to investigate whether organ culture has an effect on AdipoR1 mediated vascular function,whether organ culture affects the expression of AdipoR1 in VSMCs,and the mechanism in this process.Methods:An organ culture model in vitro was constructed after the mesenteric artery in rats were isolated and endothelial cells were removed.Vascular tone recording system(Myograph)was used to detect vascular relaxation function,Western blot and immunohistochemical staining(IHC)were used to detect the protein expression in VSMCs,and real-time fluorescent quantitative PCR(qPCR)was used to detect the mRNA expression.Results:1.Effect of organ culture on AdipoR1-mediated vascular function in VSMCs:Myograph results showed that organ culture enhanced the AdipoR1-mediated vasodilation in time-dependent manners.2.Expression of AdipoR1 in VSMCs after organ culture:Western blot and IHC results showed that protein expression of AdipoR1 was significantly enhanced after organ culture.QPCR results showed that the mRNA expression of AdipoR1 in the OC group was more than that in the Fr group in timedependent manners.3.The expression of AdipoR1 was affected by the transcriptional or translational manners during organ culture:Actinomycin D(AcD)was added to inhibit transcription.Myograph results showed that AdipoR1-mediated vasodilation in the AcD group was significantly restrained,qPCR results showed that AcD inhibited the expression of AdipoR1 mRNA.Cycloheximide(CHX)was added to inhibit protein translation during organ culture.Western blot results and semi-quantitative analysis showed that CHX did not affect the protein expression of AdipoR1.In IHC results and semi-quantitative experiments,there was no significant difference in the protein expression of AdipoR1 between the OC group and the CHX group.4.The mechanism of AdipoR1 regulation during organ culture:Myograph results showed that inhibitors of intracellular signal transduction pathway,MAPK or NF-κB pathway,did not affect AdipoR1-mediated vasodilation.Western blot results and semi-quantitative analysis showed no significant between each inhibitor group and the vehicle group.PCR results showed that the expression of AdipoR1 mRNA was no significant.Conclusion:Organ culture of rat superior mesenteric artery in vitro enhanced AdipoR1-mediated vasodilation and up-regulated AdipoR1 mRNA and protein expression in VSMCs,which were independent of activation of MAPK ERK1/2 and p38 pathways and NF-κB pathway.