Study On Regulation Mechanism Of Warming Yang, Supplementing Heart, Promoting Blood Circulation And Resolving Phlegm Therapy On NF-κB Signal Transduction Pathway In Myocardial Ischemic Reperfusion Injury

Recently,it has been reported that inflammatory response is one of the main mechanisms of myocardial ischemic reperfusion injury(MI/RI). A large amount of inflammatory cytokines are released and play an important role in the progression of MI/RI.NF-κB signal transduction pathway participates in vital physiological processes,such as cell proliferation,inflammation and apoptosis.Therefore,this research regards the inflammatory response of MI/RI as research main track and mainly focuses on NF-κB signal transduction pathway.The study used Wenxin compound as the intervening method to reveal anti-inflammatory effects and treatment target of warming yang and supplementingheart;promoting blood circulation and resolving phlegm(ab.as WSPR) on cellular and molecular level.Moreover,the research provides experimental evidence for optimizing treatment strategies and improving clinical effects.Objective:To investigate the intervention effect of WSPR on NF-κB signal transduction pathway of MI/RI,showing the anti-inflammatory treatment target andmechanism of WSPR.Method:MI/RI rat models were established by ligating the anterior branch of the left coronary artery and the myocardial ultrastructure at the ischemia region was observed under the transmission electron microscope.The content of CK and LDH in blood serum were measured with continuous monitoring using automatic blood chemistry analyzer.Besides, NIK,IKKβand IκBαprotein expression of myocardial tissues in rats were detected by western-blot;the gene and protein expression of NF-κBp65 were detected by immunohistochemistry and RT-PCR respectively.Moreover,the levels of serum IL-1β,IL-6 and IL-10 were measured by ELISA,and TNF-α, ICAM-1 expressions in myocardium were determined with immunohistochemistry.Results:①Wenxin compound could significantly improve myocardial ultrastructural changes and inhibit the release of LDH-L and CK,decrease the incidence rate of reperfusion arrhythmias;②Wenxin compound was able to inhibit the protein expression of NIK,IKKβand enhanced the expression of IKBα,the difference is significant compared to the model group(P＜0.01);③Compared with model group,the gene and protein expression and the nuclear translocation of NF-κBp65 in the Wenxin compound were obviously inhibited(P＜0.01);④Wenxin compound could significantly decrease the contents of NF-κB downstream inflammatory factors such as IL-1β,IL-6 and TNF-α,ICAM-1(P＜0.01) and simultaneously strengthen the expression of IL-10.The differences of all the indicators were significant comparing to the model group(P＜0.01) and the effect of low dose is superior to high dose.Conclusion:WSPR has cardioprotective effects against MI/RI in rats possibly by inhibiting the expression of Ck and LDH,and reducing arrhythmia and the injury of the myocardial ultrastructure.WSPR could protect myocardium from inflammatory damage during MI/RI,its mechanism might restrain the activation of upstream kinase of NF-κB signal transduction pathway and then inhibiting nuclear translocation of NF-κB,inhibiting the release of the inflammatory factors and promoting the expression of anti-inflammatory cytokines.