| Background:The incidence of atherosclerosis is high,and acute clinical events caused by it seriously endanger human health.The essence is the inflammatory response of the arteries,and cell adhesion molecules throughout the entire process of AS,mediate the adhesion of inflammatory cells,and affect the progress of the disease.Lipopolysaccharide is the main component of the outer membrane of Gram-negative bacteria,and is often used to establish a variety of inflammation models.Among them,the cell signal transduction pathway in the "two hit" rat inflammation model is unknown.In view of the important role of CAMs in AS,this subject will investigate this issue.Methods:In this experiment,a rat model of acute inflammation was first constructed.Six SD rats weighing about 300 g were randomly divided into two groups.The experimental group was injected with LPS(5 mg/kg,n=3)intraperitoneally,and the control group was injected.The volume of normal saline(n=3)was injected continuously for two days.After the first injection,the animals were fasted overnight and had free access to water.12 hours after the second injection,the animals were killed by decapitation after CO2 asphyxiation,and the thoracic aorta was removed with Triton X-100 to endothelium.High-throughput TMT-labeled proteomics was used to screen out differential proteins under different processing conditions.To verify the omics results,we applied model rat thoracic aorta tissue(with endothelium removed)and cultured rat thoracic aortic smooth muscle cell line A7R5 cells in vitro,focusing on the activation of cell adhesion molecules and related signaling pathways in thoracic aortic smooth muscle induced by LP S.Results:The quality control results of TMT-labeled proteomics of smooth muscle tissue of rat thoracic aorta without endothelial were qualified,and the sample had good repeatability.5176 differential proteins were identified by experiments,and 1.5 times was selected as the multiple of difference.It was found that 410 proteins were up-regulated and 98 proteins were down-regulated.The cluster analysis results of KEGG pathway enrichment showed that the changes of JAK-STAT and CAMs induced by LPS were particularly obvious,and NF-κB also changed to some extent.The tissue Western blot results confirmed that LPS can activate p-STAT3,VCAM and ICAM,and the difference was statistically significant.The results of smooth muscle cell experiments showed that LPS can increase the phosphorylation level of p-p65 and p-STAT3,and the expression of VCAM and ICAM also increased.Inhibition of JAK-STAT with the inhibitor JANEX-1 was found to reduce the expression of VCAM,ICAM and p-p65.Conclusion:The expression of CAMs was increased in the smooth muscle of thoracic aorta in rats with two LPS attacks,so VCAM and ICAM are expected to be used as diagnostic and stratification indicators for AS.The increase of CAMs may be related to JAK-STAT and NF-κB,suggesting that the inflammatory response of AS can be reduced by inhibiting the activation of JAK-STAT and NF-κB.And the inhibitor JANEX-1 can be used in the clinic as an anti-inflammatory drug.The modification differences in the inflammation model suggest that inflammation is related to modification,which provides a new perspective for the study of AS and can be used as the future research direction of AS Therefor e,the study of these signaling pathways is of certain reference value for the diagnosis and treatment of chronic inflammation AS. |
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