The Effect Of Angiotensin-? On NLRP3 Inflammasome In H9C2 Cells

Background:Chronic heart failure is the terminal stage of various cardiovascular diseases.Domestic and foreign researchers generally believe that ventricular remodeling is the main physiopathologic mechanism of chronic heart failure.Angiotensin-II(AngII)can induce ventricular remodeling by activating the neuroendocrine system.However,a growing number of evidence suggests that AngII can also induce ventricular remodeling by activating interleukin-6(IL-6)and transforming growth factor-?(TNF-?)or other inflammatory cytokines.NLRP3 inflammatory body and its downstream products are involved in myocarditis caused by various etiologies,and further lead to ventricular remodeling.However,there are few reports about whether AngII can directly induce inflammation of cardiomyocytes through NLRP3 inflammatory bodies or not.This study aims at analyzing the impacts of AngII on heart failure through inflammation.Objective:To explore the effects of angiotensin-II on NLRP3 inflammation in H9C2 cells.Methods:All experiments were performed by using of the H9C2 cells,which were cultured in vitro and treated with 10-6mol/l of AngII.The cells were collected at varying time points(0h?1h?3h?6h?12h).Expression levels of NLRP3,ASC,Caspase-1,IL-1? mRNA were detected with quantitative real-time PCR,expression levels of NLRP3,ASC,Caspase-1,IL-1? protein were detected by western blot,and the content of inflammatory factors in supernatant detected through ELISA.Results:1.With the treatment of AngII(10-6mol/l)for varying lengths of time(0h?1h?3h?6h?12h),the expression of mRNA levels of NLRP3,ASC,Caspase-1and IL-1? were significantly higher compared with 0h group(P<0.05),except for that of the Caspase-1 in 1h group,which showed no difference compared with 0h group.There existed significant differences when respectively comparing the mRNA levels of NLRP3,ASC,Caspase-1 and IL-1? in one group with that in any other group among 1h,3h,6h and 12 h groups(P<0.05),except for in 1h group and 3h group(P>0.05),between which there is no difference in the mRNA levels of NLRP3.And the remaining pairwise comparison of different groups showed significant difference(P<0.05).2.With the treatment of AngII(10-6mol/l)for varying lengths of time(0h?1h?3h?6h?12h),the expression of protein levels of NLRP3,ASC,Caspase-1and IL-1? were significantly higher than those in 0h group(P<0.05).And there were significant differences between different time groups(P<0.05).3.With the treatment of AngII(10-6mol/l)for varying lengths of time(0h?1h?3h?6h?12h),the content of inflammatory factors IL-1? in the supernatant were significantly higher than those in the 0h group(P<0.05).And there were significant difference between different time groups(P<0.05).Conclusion:1.With the treatment of AngII(10-6mol/l)for varying lengths of time(0h?1h ? 3h ? 6h ? 12h),H9C2 cells grew in well and there was no obvious abnormality in H9C2 cells morphology.2.Treatment to H9C2 cells with AngII could give rise to the mRNA andprotein expression levels of NLRP3,ASC,Caspase-1 and IL-1?,and increase the content of IL-1? in the supernatant.3.Angiotensin-II could induce inflammation of cardiomyocytes by activating NLRP3 inflammatory bodies,and this process is associated with angiotensin-II leading to heart failure,and provides theoretical basis for clinical treatment of cardiac dysfunction and myocardial remodeling caused by myocarditis.