| Background and objective:Atherosclerosis(atherosclerosis,AS)is a chronic inflammatory vascular disease mainly involved the large and middle arteries,where usually occurs at the site of endothelial cell damage and blood flow disorders.Its complications are acute and can threaten life in severe conditions,which bring great harm to human health.Studies have shown that inflammation is closely related to the occurrence and development of AS.Inflammatory cells and inflammatory factors participate in all stages of the disease,from early endothelial damage to plaque formation and even plaque rupture.When the blood vessels continue to be stimulated by some factors such as inflammation,adherens junction between endothelial cells is weakened and thereby endothelial barrier is also damaged,which promotes the development of AS.Transmembrane 4 L Six Family Member 19(TM4SF19),TM4SF19 has been reported about obesity,but there are still no essays about TM4SF19 and AS.Therefore,the purpose of this study is to explore the related mechanisms of TM4SF19 in the pathological process of AS,observe whether inflammation has an impact onTM4SF19,and whether LPS can affect adherens junction through TM4SF19 and other pathways.In this way,we supplement adherens junction related molecular mechanisms in AS and provide new target for prevention and treatment of AS.Methods:Human umbilical vein endothelial cells(HUVECs)were cultured in vitro.Quantitative real-time polymerase chain reaction(qRT-PCR)and Western Blot were used to detect TM4SF19 and VE-cadherin expression in LPS different concentrations and time treatments.Small interference fragments and plasmid transfection experiments,qRT-PCR,Western Blot,and immunofluorescence were used to investigate the impact of TM4SF19 on VE-cadherin,molecular of adherens junction and its related mechanisms.Then 5 pairs of normal arterial endometrial tissue and human atherosclerotic plaque tissue were collected.The location and expression of TM4SF19 in plaque tissues were detected by immunohistochemistry(IHC).At last,peripheral blood blood samples were collected from healthy people and clinically AS-diagnosed patients.The serum of TM4SF19 expression level and ROC curve were used to analyze the diagnostic value of TM4SF19 in AS by ELISA.Results:1.LPS can increase the expression level of TM4SF19 in HUVECs in a concentrationand time-gradient-dependent manner.As the LPS concentration increases or the processing time prolongs,TM4SF19 expression in the treatment group gradually increases,and the difference is statistically significant(P<0.05).2.When HUVECs were treated with LPS at different concentrations and times,the mRNA and protein levels of VE-cadherin were down-regulated in a concentrationand time-dependent manner.3.HUVECs were transfected with small interfering fragments and recombinant plasmids,knockdown of TM4SF19 could up-regulate the expression of VE-cadherin in HUVECs,while over-expression of TM4SF19 could markedly reduce VE-cadherin mRNA and protein expression levels.4.Overexpression of TM4SF19 on the basis of LPS stimulation can further inhibit VE-cadherin expression and disrupt HUVEC adhesion connections,while knocking down TM4SF19 partially reversed the weakening of HUVECs adhesion connections induced by LPS.5.Immunohistochemistry results showed that TM4SF19 was significantly higher expressed in human atheromatous tissue than in normal human arterial intimal tissue.6.ELISA results showed that TM4SF19 was significantly increased in the serum of AS patients,and ROC curve analysis revealed that TM4SF19 had valuable diagnostic performance for AS.Conclusion:1.LPS could upregulate TM4SF19 expression and downregulate VE-cadherin expression in HUVECs in a concentration dependent manner.2.TM4SF19 can mediate LPS to inhibit VE-cadherin expression and thus weaken the adhesion and connection between vascular endothelial cells.3.TM4SF19 is highly expressed in atherosclerotic plaque tissue and peripheral blood samples of AS patients,and has good diagnostic performance for AS. |
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