Effect Of Interleukin-12p35 In Inflammatory Response And Autophagyon On Angiotensin Ⅱ-induced Myocardial Hypertrophy Mice

ObjectiveThis experiment use AngⅡ-induced hypertensive myocardial hypertrophy model in wild-type mice and interleukin-12p35(IL-12p35)knockout mice on C57BL/6J background to study the expression of IL-12p35 during the development of cardiac hypertrophy,to investigate the role of IL-12p35 gene in the regulation of inflammation and autophagy during cardiac hypertrophy.This provides a new therapeutic target and experimental basis for clinical treatment of angiotensin II(AngⅡ)-induced hypertensive myocardial hypertrophy.Methods1.Animals:We used C57BL/6J male mice aged 10-12 weeks,were divided into two groups,A: Negative control group(n=8),B: Positive control group(n=8).IL-12p35 knockout mice(C57BL/6J background)aged 10-12 weeks were divided into two groups,C: experimental group(n=8),D:Conditional control group(n=8).2.Blood pressure measured:Blood pressure and heart rate was measured by tail-cuff system(BP-98 A,Softron,Japan)evey week.3.Echocardiography:Data acquisition was performed using a small animal ultrasound system(Visual Sonics Vevo 2100).4.Cardiac hypertrophy:The expression of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP)and Collagen I were detected by Real-time quantitative polymerase chain reaction5.heart weight/body weight observed myocardial hypertrophy.6.Myocardial fibrosis:Myocardial tissue fibrosis was examined using Masson’s trichrome staining.7.Inflammatory infiltration:To test CD3,CD68 expression by immunohistochemical and hematoxylin-eosin staining.8.Autophagy:Microtubule-associated protein 1 light chain 3(LC3),p62 expression was detected by Western blot and immunohistochemical staining.9.Expression of IL-12p35:IL-12p35 expression was detected by Real-time quantitative polymerase chain reaction.10.Signal pathway:Western Blot detection of JAK2/STAT4 channel phosphorylation.11.Statistics:Use SPSS18.0 software for statistical analysis of data,all data are expressed as mean±standard deviation.t-test was used to compare the two groups,and one-way ANOVA was used to compare the multiple organizations,P<0.05 was considered statistically significant,P<0.05 was statistically significant,P<0.01 was significantiy statistical difference,and P<0.001 was extremely significant statistical difference.ResultsThere was no statistical difference in basic blood pressure between the 4 groups before AngⅡ perfusion,and the blood pressure increased significantly on the 3th day after Angll perfusion,and reached the peak at 3th week,lasting to 4th week.The results of ultrasound and ANP mRNA,BNP mRNA,Collagen I mRNA showed that myocardial hypertrophy was successfully modeled.SBP and HW/BW in IL-12p35-KO+AngⅡ group were significantly higher than WT+AngⅡ group,and there was statistical significance.HE and Masson staining reflected the myocardial fibrosis,inflammatory cell infiltration,myocardial cell hypertrophy,disordered arrangement,nuclear deformity and vacuolation are more significant,IL-12p35-KO+AngⅡ group was significantly higher than WT+AngⅡ group,and it was statistically significant.AngⅡ perfusion increased the expression of CD3 and CD68.IL-12p35 deficiency enhanced autophagy and promoted phosphorylation of the JAK2/STAT4 signaling pathway,whereas AngⅡ perfusion exasperated autophagy and phosphorylation of the JAK2/STAT4 signaling pathway.After AngⅡ perfusion,the expression of IL-12p35 mRNA in the WT+AngⅡ group was significantly higher than that of WT group.ConclusionsIn the model of myocardial hypertrophy induced by AngⅡ,Studies have shown that IL-12p35 deficiency can promote higher blood pressure,increase HW/BW,aggravate myocardial fibrosis,inflammatory cell infiltration,myocardial cell hypertrophy,disordered arrangement,nuclear deformity,vacuolation and autophagy levels,and also activate the phosphorylation of the JAK2/STAT4 signaling pathway.The expression of IL-12p35 mRNA in the WT+AngⅡ group was significantly higher than WT group.It suggested that IL-12p35 not only participated in the pathological process of myocardial hypertrophy induced by AngⅡ,but also had the effect of inhibiting inflammation and regulating autophagy,and these biological effects may be mediated by JAK2/STAT4 signaling pathway.This showed that IL-12p35 may become a new target for the prevention and treatment of myocardial hypertrophy...