Oleoylethanolamide Modulates Maturation Of Dendritic Cells To Reduce Arterial Intimal Hyperplasia And Its Mechanisms

Objective: The initiation and progression of atherosclerosis is primarily driven by the recruitment of inflammatory cells and the secretion of cytokines.The purpose of this study was to investigate whether the compound oleoylethanolamide(OEA)inhibits the maturation of dendritic cell(DC)and inhibits arterial intimal hyperplasia to reduce atherosclerosis.OEA has been found to be a bioactive lipid that reduces cerebral infarction,cerebral cortical apoptosis,and alleviation of intimal hyperplasia after cerebral ischemia and reperfusion.However,whether OEA partially inhibits intimal hyperplasia by regulating DC immune maturation remains to be identified.Methods: In this study,we used in vitro and in vivo experiments to demonstrate the effect of OEA on DC maturation.In vitro experiments,6-8 weeks old male C57BL/6 mice were used to obtain bone marrow-derived DC cultured in 6-well plates.The groups were divided into control group,experimental group(LPS 200 ng/ml),and drug-administered group(OEA 20,40,80 u M),after 7 days of culture,the DC surface maturation markers CD80,CD86,MHCII were examined by flow cytometry analysis,DC migration ability was tested by cell migration assay,and T Cell proliferation was determined in co-cultured DC and T cell systems.,and production of cytokines IL-6,IL-12,TGF-? were measured by using QPCR.To elucidate whether OEA affects the TLR4 / NF-?B pathway,we used Western blotting to determine the protein level of TLR4 and the phosphorylation level of protein p65.At the same time,it was investigated whether the inhibition of DC maturation by OEA through its receptor transient receptor potential vanilloid-1(TRPV-1)and its underlying mechanism.TRPV1 antagonist CPZ was added to the experiment.In addition,to investigate the role of protein kinase AMPK in OEA-induced inhibition of TLR4/NF-?B signaling pathway through TRPV1 activation,AMPK antagonist compound C(Compound C,CC)was added.In vivo,6-8 weeks old male C57 BL / 6 mice,10 in each group,OEA group dose of 10 mg / kg,20 mg / kg and 40 mg / kg preprotected for 3 days,intraperitoneal injection of LPS 2.5ug induced spleen DC maturation.In the balloon injury model,male SD rats of about 250-280 g were used,and 10 rats in each group were divided into sham operation group,BI group,OEA group,and the left common carotid artery(CCA)was damaged 3 times with a 2F balloon catheter.OEA intervention was given before balloon injury and 4 days after injury.Results: OEA was observed to inhibit DC maturation in vitro.The concentration of Ca2+ in the DC increased after OEA treatment,and the effect of OEA on MHCII in the spleen of mice treated with CPZ was eliminated in LPS induced spleen DC maturation in mice.The TLR4/NF-?B pathway is a classical pathway by which LPS induces DC maturation.The results showed an increase in LPS stimulation,and OEA treatment significantly decreased the protein levels of TLR4 and p-p65.At the same time,the CPZ group significantly impeded the reduction of TLR4 / NF-?B by OEA.The results showed that blocking AMPK had no effect on reducing TLR4 in the OEA treated group,but effectively reversed OEA inhibition of NF-?B levels.This indicates that OEA down regulates TLR4/NF-?B by activating TRPV1 and activating AMPK.Taken together,these results suggest that OEA inhibits DC maturation by activating TRPV1 and AMPK,which may help to reduce intimal hyperplasia induced by balloon injury.Correspondingly,in the LPS induced DC maturation mouse model,OEA reduced the maturation markers on spleen DC,and OEA reduced the accumulation of DC in the early neointimal induced by balloon injury in rats,two animal models illustrating OEA The inhibition of DC maturation is consistent,which helps to alleviate neointimal hyperplasia through OEA.These findings add to our understanding of this endogenous lipid OEA.Conclusion:(1)OEA can inhibit DC maturation and reduce immune inflammatory response;(2)OEA can inhibit the number of DC,reduce the intimal hyperplasia caused by balloon injury,and has significant vascular protection;(3)OEA may be Inhibition of DC maturation is achieved by activating the TRPV-1 receptor to regulate the TLR4/NF-?B signaling pathway.