The Study Of NLRP3 Inhibitor MCC950 In Vascular Remodeling

Objective:when the internal and external environment changes,blood vessels will make a series of structural and functional changes to adapt,this adaptation process is called vascular remodeling.Vascular smooth muscle cell dedifferentiation is the core of vascular pathology,such as atherosclerosis,restenosis and aneurysm.NLRP3 inflammato ry bodies are associated with the occurrence and progression of various diseases,including metabolic disorders,multiple sclerosis,inflammatory bowel disease,and other autoimmune and autoinflammatory diseases.Recent studies have shown that NLRP3 gene d eletion can reduce Ang II-induced inflammation,VSMC phenotypic transformation and proliferation,as well as Ang II-induced hypertension and vascular remodeling.However,there are few studies on the mechanism of vascular remodeling after acute vascular in jury,and the mechanism is unknown.To investigate the inhibitory effect of NLRP3 inhibitor MCC950 on the proliferation,migration and phenotypic transformation of smooth muscle cells induced by Angiotensin-converting enzyme(Ang Ⅱ),to study the role of NL RP3 in vascular remodeling induced by arterial ligation,to explore the mechanism of vascular remodeling caused by acute injury,and to find a new target for inhibiting vascular injury,we designed and carried out this experiment.Methods:In the part of cell experiment,3-5 generations of human aortic smooth muscle cell lines were selected for smooth muscle cells.The VSMC was digested with trypsin,then inoculated into a 60 mm petri dish and cultured in a humid environment of 37 °C,95% air and 5%CO2.Be fore the experiment,smooth muscle cells were cultured in serum-free medium(SMGS)for 24 hours to achieve rest,and then incubated with Ang Ⅱ(1μmol/L)for 12 hours to induce inflammation and proliferation.According to the experimental conditions,they were divided into Ang Ⅱ group,Ang+MCC950 group,MCC950 group and PBS control group.Ang II+MCC950 group was pretreated with MCC950(1μmol/L)before Ang II induction the proliferation of smooth muscle cells.The proliferation of smooth muscle cells was dete cted by CCK-8 proliferation test and EDU proliferation test.The changes of migration ability of smooth muscle cells were detected by scratch test and Transwell test.Western blot and q RT-PCR were used to detect the protein expression of contractile phenot ype and proliferative phenotype in vascular smooth muscle cells.In the animal experiment,24 male C57 BL /6 mice weighing 20-30g(9 to 10 weeks)were selected to establish the model of carotid artery ligation.According to the ligation of common carotid artery and injection of MCC950,the animals were randomly divided into four groups: simple carotid artery ligation group,MCC950(10mg/kg)injection and carotid artery ligation group,MCC950(10mg/kg)injection g roup and normal saline injection control group.Carotid immune section was used to detect vascular proliferation,and Western Blot and q RT-PCR were used to detect the expression of contractile phenotype and proliferative phenotype related proteins of vascu lar smooth muscle cells.Results:1.AngⅡ can induce proliferation,migration and phenotypic transformation of vascular smooth muscle cells.2.The intervention of MCC950 can inhibit the proliferation,migration and phenotypic transformation of vascular smooth muscle cells induced by Ang Ⅱ,but MCC950 has no inducing or inhibitory effect on normal vascular smooth muscle cells.3.After ligation of carotid artery,intima and media proliferated obviously,and phenotypic transformation of smooth muscle cells occurred.4.MCC950 can inhibit vascular r emodeling and phenotypic transformation of smooth muscle cells after carotid artery ligation.Conclusion:1.MCC950,a specific inhibitor of NLRP3 inflammatory bodies,can inhibit the proliferation,migration and phenotypic transformation of smooth muscle cells induced by Ang Ⅱ,indicating that NLRP3 plays an important role in Ang II-induced inflammation,phenotypic transformation,vascular smooth muscle cell proliferation and vascular remodeling,which provides an idea for the prevention and reversal of vascular remodeling at cellular level.2.MCC950 can inhibit the proliferation,migration and phenotypic transformation of smooth muscle cells after carotid artery injury,suggesting that NLRP3 plays an important role in vascular remodeling after acute caroti d artery injury,which provides a new target for inhibiting vascular remodeling after acute carotid artery injury.