The Effects Of Anti-inflammation Of Propane-2-sulfonic Acid Octadec-9-enyl-amide And Oleoylethanolamide

Background:Atherosclerosis(AS)is a common pathophysiology basis of many cardiovascular and cerebrovascular diseases and is one of the most common cardiovascular and cerebrovascular diseases.AS is a disease that begins with an arterial intima,and its pathogenesis mechanism has many different opinions.High blood pressure,high cholesterol,smoking,diabetes and obesity are its risk factors.In recent years,the establishment of the theory of inflammation indicates the direction for the AS study.N-oleoyl ethanolamine(OEA)is a naturally occurring in vivo fatty acid alcohol amine compound,which is one of the agonists of peroxisome proliferator-activated receptor-a(PPARa).Our former study has found that OEA can inhibit the expression of TLR4 by activating PPARa and inhibit the inflammation of THP-1 response to the stimulation of LPS.But the specific anti-inflammatory mechanism of OEA is not clear.Propane-2-sulfonic acid octadec-9-enyl-amide(N15)is a self-designed OEA analogues compound with PPARa agonism.The results of early experiments showed that N15 could inhibit the expression of VCAM-1 and ICAM-1 in vascular endothelial cells,which was of great significance to the early prevention and treatment of atherosclerosis.But its specific anti-inflammatory mechanism is not clear.So the anti-inflammatory effects of N15 and OEA will be studied in this paper.Aims:To study the anti-inflammatory mechanism of N15 and OEA.Methods:1.To investigate the anti-inflammatory mechanism of N15:THP-1 cells were treated with different concentrations of N15 pre-protection and then induced by LPS.The expression of TLR4 and NF-?B related signaling molecules will be detected by Western Blot and real-time PCR.2.To investigate the anti-inflammatory mechanism of OEA:(1)In vivo experiments:Atherosclerosis model was established by injecting Angiotensin ? into APOE-/-knockout mice.The 2 month old male APOE-/-gene knockout mice were randomly divided into Control group,Model group,OEA30 group(OEA-30mg/kg),After 30 days of administration,blood vessels were taken which were observed and detected the expression of inflammatory proteins using Western Blot.(2)In vitro experiments:THP-1 cells treated with different concentrations of OEA preconditioning after were adhered with PMA and then induced inflammatory response of THP-1 cells with LPS.The migration of THP-1 cells was observed by Transwell chamber experiment and the expression of inflammatory protein was detected by Western Blot and real-time PCR.Results:1.N15 inhibited the activation of TLR4,NF-?B and the phosphorylation of STAT3 in LPS-induced THP-1 cells.After silencing the PPAR-gene,the inhibitory effect of N15 on TLR4,NF-?B and STAT3 phosphorylation was decreased.2.OEA can reduce the proliferation of aortic intima and the migration of THP-1 monocytes to macrophages.It can also enhance the phosphorylation of AMPK and inhibit the expression of phosphorylated STAT3.After silencing AMPK and PPAR-a,the inhibition effect of OEA on the migration of THP-1 monocytes to macrophages was decreased,and also the effect of OEA on enhanced phosphorylation of AMPK and inhibited the phosphorylation of STAT3 was decreased.Conclusion:1.In LPS-induced THP-1 cells,N15 exerts anti-inflammatory effects by inhibiting TLR4 and downstream NF-?B and STAT3 signaling pathways.2.OEA can play an important role in anti-inflammation and anti-atherosclerosis by reducing the migration of monocytes to macrophages and the phosphorylation of STAT3 and increasing the expression of PPARa and the phosphorylation of AMPK.