Protein Kinase TBK1/IKK-? Inhibitor Amlexanox Improves Cardiac Function In Mice After Acute Myocardial Infarction

BackgroundRecent studies have shown that innate immunity involving non-classical IKK-related protein kinases TBK1 and IKK-? play an important role in the development of acute myocardial infarction(AMI).Clinically,Amlexanox is a primary drug that is used to treat asthma,allergic rhinitis,and recurrent oral ulcers.Previous studies have shown that Amlexanox can reduce chronic inflammation,increase insulin sensitivity,improve energy metabolism,and alleviate liver steatosis by blocking TBK1 and IKK-?inflammatory signaling pathways.However,whether Amlexanox can affect the occurrence and development of AMI,and related mechanisms have not been reported.In this paper,we explored whether the effect of Amlexanox affects cardiac function after AMI in mice,and its possible mechanisms.Objectives1.To study the effects of protein kinase TBK1 and IKK-? inhibitor Amlexanox on cardiac function in mice post-AMI;2.To explore the specific mechanism of protein kinase TBK1 and IKK-? inhibitor Amlexanox affecting cardiac function in mice post-AMI,and provide a new possibility for clinical improvement of prognosis in patients with AMI.MethodsIn vivo study1.Establish acute myocardial infarction(AMI)model in mice Mice were divided into 4 groups randomly:sham+DMSO group,sham+Amlexanox group,AMI+DMSO group,AMI+Amlexanox group,of which about 20 mice per group.2.About 12 hours after the surgery,the sham+Amlexanox group and the AMI+Amlexanox group were given a dose of 25 mg/kg for intraperitoneal injection of Amlexanox to the mice,once daily for 7 days,sham+DMSO group and AMI+DMSO group was given an equal amount of DMSO as a control.3.Cardiac function in 3 days and 7 days after operation in each group of mice was evaluated with echocardiography.4.The mice's body weight before operation and body weight,heart weight and tibia length were measured 7 days after operation in each group of mice.5.RT-PCR was used to detect the mRNA transcription levels of IFNb1,IFIT1,IFIT3,IRF7,CXCL10,ISG15,Collal,Col3al,ANP,BNP,Myh7 and other genes in myocardium on the 7th day after operation.6.H&E staining was used to detect the level of inflammatory infiltration of myocardium in 7 days after operation in each group of mice.7.Sirius red staining and Masson staining were used to determine myocardium fibrosis levels in 7 days after operation in each group of mice;8.Immunohistochemical staining was performed to assess the expression of MOMA2.CD31 and other proteins in myocardial tissue of mice in each group.9.Western blotting was was performed to assess the expression levels of TBK1,pTBK1,IRF3,pIRF3,Bcl-2,Caspase-3,BAX,Collagen I,Collagen III,?-Tubulin in myocardial tissue of mice in each group.Results1.Amlexanox improved left ventricular ejection fraction(LVEF)and short axis shortening rate(FS),and reduced myocardial pathological remodeling in mice post-AMI.2.Amlexanox reduced inflammatory cells infiltration and the expression of pro-inflammatory cytokines in mice post-AMI.3.Amlexanox promoted the myocardium fiber repair in the infarcted area of mice post-AMI,meanwhile reduced myocardium fibrosis in non-infarcted areas.4.Amlexanox accelerated myocardial tissue angiogenesis in mice post-AMI.5.Amlexanox alleviated myocardial apoptosis in mice post-AMI.Conclusions1.Protein kinase TBK1 and IKK-s inhibitor Amlexanox can improve cardiac function in mice post-AMI;2.Protein kinase TBK1 and IKK-s inhibitor Amlexanox can reduce myocardial inflammation after AMI in mice,promote myocardial fiber repair in infarcted area,accelerate myocardial angiogenesis,and alleviate myocardial apoptosis.