Huoxin Pill Against Acute Myocardial Ischemia Inflammatory Injury Through The Inhibition Of TLR4/NF-?B Signaling Pathway

Purposes:Animal experiments were conducted to investigate the protective effect of Huoxin pill(HXP)on ISO-induced acute myocardial ischemia(AMI)rats' heart injury,and to explore the effect of Huoxin pill(HXP)on cardiac inflammation and its potential mechanism based on the TLR4/NF-?B signaling pathway,and a series of pathological processes related to AMI induced cardiac inflammation.To further clarify the action mechanism of Huoxin pill(HXP)to improve cardiac damage caused by acute myocardial ischemia,and to provide further experimental basis for clinical Huoxin pill(HXP)to treat acute myocardial ischemia and protect target organ damage.Methods:1.The rats were randomly divided into 4 groups according to the initial body weight,and 8rats in each group: Sham group,Acute myocardial ischemia group(AMI),HXP-L group and HXP-H group.Rats in HXP-L and HXP-H groups were administered with 3 mg/kg/d and 9mg/kg/d of HXP for 10 days,respectively,rats in Sham and AMI groups were received an equivalent volume of distilled water.At the 9th and 10 th day,the rats of AMI,HXP-L and HXP-H groups were injected subcutaneously with isoproterenol(8km/kg),while the rats of Sham group were injected with an equivalent volume of saline1.Changes in ST segment were detected by electrocardiogram(ECG).2.Echocardiography detected cardiac function.3.The serum levels of CTnT,CK-MB,SOD,MDA,IL-6 and TNF-a of rats in each group were determined using ELISA analysis.4.The heart weight was calculated and HE staining was performed to understand the pathological changes of cardiac tissues.5.Immunohistochemistry(IHC)detected the protein expression of Mac-2,interleukin-6(IL-6),tumor necrosis factor alpha(TNF-?),TLR4,NF-?B and p65 in cardiac tissues Results:1.With the treatment of HXP,the ST-segment elevation was significantly attenuated in the HXP-L and HXP-H group while compared with the AMI group.2.compared with Sham group,the end-diastolic volume of rats AMI group were significantly increased,which obviously attenuated after 9 mg/kg of HXP treatment.3.As two major indicators of myocardial injury marker enzymes,CTnT and CK-MB levels were significantly increased in serum of AMI rats,which were dramatically reduced after HXP treatment;ISO injection obviously increased the levels of MDA in serum of AMI rats,which was alleviated after HXP treatment;In contrast,ISO injection obviously decreased the activities of SOD,which were increased after 9 mg/kg of HXP treatment.the serum levels of IL-6,TNF-? were obviously increased after ISO injection,which were obviously reduced after HXP treatment.4.Heart weight was observedas higher in the AMI group rats than sham group rats.after administration of HXP(9mg/kg),the heart weight was significantly decreased.H&E staining showed that ISO-treated rats exhibited apparent myocardial cell edema,degeneration,transverse striations loss,and increased inflammatory cell infiltration,which were significantly attenuated after administration of HXP.5.Immunohistochemistry analsyis revealed that the protein expression of Mac2,IL-6,TNF-a,TLR4 and p65 in cardiac tissues were significantly increased in AMI group.On the contrary,HXP treatment obviously decreased the protein expression of Mac2,IL-6,TNF-a,TLR4 and p65 in cardiac tissues.Conclusions:1.Huoxin pill(HXP)could decrease the ST elevation induced by AMI.2.Huoxin pill(HXP)could improve cardiac function;3.Huoxin pill(HXP)could improve myocardial enzyme and oxidative stress injury caused by acute myocardial ischemia;reduce cardiac weight and improve cardiac pathological morphology.4.Huoxin pill(HXP)could reduce the release of Mac-2,IL-6,TNF-a inflammatory factor and have a protective effect on heart damage.5.Huoxin Pill(HXP)could improve cardiac inflammation caused by acute myocardial ischemia by regulating the expression of TLR4 and p65 in TLR4/NF-?B signaling pathway.