Exploring The Effects And Mechanisms Of Wenyang-Yixin Therapy In Coronary Heart Disease With Depression Based Into NLRP3

Coronary heart disease(CHD)with depression is a group of clinical syndromes with unknown pathogenesis and multi-system co-morbidities,with chest tightness,chest pain,depressed mood and pessimistic depression as the main manifestations,which seriously affects patients’ physical and mental health.It is a complex disease with high incidence,low diagnosis rate,lack of effective prevention and treatment methods,and prone to unexpected events,which are the problems and keys of prevention and treatment.Coronary heart disease with depression interact with each other and have intricate clinical manifestations.Routine treatment with coronary heart disease drugs combined with antidepressant and sedative drugs can alleviate symptoms to some extent,however,some antidepressant drugs can easily induce myocardial ischemia and arrhythmia,which affect the prognosis of coronary heart disease.Through more than 30 years of clinical practice and research,Professor Cao Hongxin believes that Yang deficiency and phlegm stasis is the core pathogenesis of coronary heart disease,and proposes Wenyang-Yixin therapy to treat coronary heart disease,and established the representative prescription "Wenxinfang" as the basic method for treating coronary heart disease.In recent years,the number of patients with coronary heart disease combined with depression has gradually increased.According to the symptoms,the combination of nervine and tranquilizer on the basis of Wenxinfang can warm yang,tonify qi and benefit the heart,invigorate blood,dissolve phlegm and open the veins,relieve depression and nervine and tranquilize the mind,which can obviously improve the physical and mental symptoms of patients with coronary heart disease and depression.Our group’s previous research confirmed that the Wenyang-Yixin therapy can up-regulate the sugar-water preference index of coronary heart disease with depression model,increase the number and time of entering the central zone in the open field experiment,reduce the immobility time in the forced swimming experiment,and significantly improve the depressive state;promote hippocampal pentraxin,dopamine,brain-derived neurotrophic factor,tyrosine kinase receptor B protein expression,and play a role in protecting hippocampal neurons and synaptic plasticity.Modern medical research has shown that inflammation affects neurological,endocrine,and immune regulation and is one of the core pathogenic mechanisms connecting in coronary heart disease with depression.Objective:In this study,we investigated the mechanism of inflammatory effects of Wengyang-Yixin therapy to regulate NLRP3 inflammatory vesicles to inhibit coronary heart disease with depression through animal experiments.1.Modeling coronary heart disease with depression using high-fat chow-fed ApoE-/-mice combined with chronic unpredictable mild stress;2.To clarify the differences of NLRP3 inflammatory related protein and mRNA expression in coronary heart disease with depression,coronary heart disease and depression mice;3.To investigate the effect of Wenyang-Yixin therapy on NLRP3 inflammatory related protein and mRNA expression in mice with coronary heart disease and depression,and to provide scientific basis for the treatment of coronary heart disease with depression by WenYang-Yixin therapy.Methods:1.ApoE-/-mice were fed high-fat chow for 12 weeks to establish a coronary heart disease model,ApoE-/-mice were fed chronic unpredictable mild stress for 4 weeks to establish a depression model,and ApoE-/-mice were fed high-fat chow combined with chronic unpredictable mild stress for 16 weeks to establish a coronary heart disease with depression model.The successfully molded mouse models of coronary heart disease with depression were randomly divided into 6 groups:coronary heart disease with depression group(equal dose of pure water),Wenxinfang high dose group(47.84g/kg/d),Wenxinfang medium dose group(23.92g/kg/d),Wenxinfang low dose group(11.96g/kg/d),western medicine group(fluoxetine hydrochloride 2.6mg/kg/d and atorvastatin calcium 1.3mg/kg/d),agonist group(intraperitoneal injection of trimethylamine N-oxide 25μg/kg+Wenxinfang 23.92g/kg/d),using C57BL/6 mice as blank group(equal dose of pure water),and the corresponding indexes were tested after 28 days of continuous gavage.The depressive behavior of mice was assessed using sugar-water preference test sugar-water consumption ratio and forced swimming test,hanging tail test immobility time;HE staining to observe the morphology of atherosclerotic lesions in mice aorta and calculate the percentage of plaque to assess the aortic lesions;immunohistochemical staining to observe the activation of microglia in CA1 area of hippocampus;TUNEL staining to assess neuronal apoptosis;IL-1β,IL-6,IL-10 and IL-18 were detected by ELISA;NLRP3,ASC,Caspase-1 and NF-κB p65 in mouse aorta and hippocampus were detected by Western Blot;NLRP3,ASC,Caspase-1,IL-1β,IL-18 and NF-κB mRNA in mouse aorta and hippocampus were detected by RT-PCR.Results:1.The percentage of sugar water consumption was significantly lower in the mice with coronary heart disease and depression group(P<0.01),and the duration of immobility was significantly increased in the forced swimming and hanging tail experiments(P<0.01);significant atherosclerotic plaque deposition was seen in the aorta,and the percentage of aortic plaque area was significantly increased(P<0.01).2.The number of hippocampal Iba-1 positive cells and TUNEL positive cells were significantly increased in the mice with coronary heart disease with depression group(P<0.01,P<0.01);serum IL-1β,IL-6,IL-18 levels were significantly increased(P<0.01)and IL-10 levels were significantly decreased(P<0.01);aortic NLRP3,ASC,Caspase-1,hippocampal NLRP3,Caspase-1,NF-κB p65 protein expression was significantly increased(P<0.01),aortic NF-κB p 65,hippocampal ASC protein expression was increased(P<0.05);aortic ASC,Caspase-1,IL-1β IL-18,hippocampal NLRP3,ASC,Caspase-1,IL-1β,IL-18 mRNA expression levels were significantly increased(P<0.01),and aortic NLRP3,NF-κB mRNA expression levels were increased(P<0.05).3.The Wenxinfang high dose group significantly increased the percentage of sugar and water consumption(P<0.01),significantly reduced the immobility time in the hanging tail experiment(P<0.01),and reduced the immobility time in the forced swimming experiment(P<0.01);significantly reduced the number of microglia activation and neuronal apoptosis in hippocampal CA1 area(P<0.01),and reduced the percentage of aortic plaque area(P<0.05);reduced aortic NLRP3,ASC,Caspase-1,NF-κBp65 and hippocampal NLRP3,Caspase-1 protein expression(P<0.01),decreased hippocampal NF-κB p65 protein expression(P<0.05);significantly decreased aortic,hippocampal NLRP3,ASC,Caspase-1,NF-κB mRNA expression levels(P<0.01),decreased aortic and hippocampal IL-1β mRNA expression levels(P<0.05).4.After using of NLRP3 inflammatory vesicle agonist trimethylamine N-oxide,aortic NLRP3,Caspase-1,NF-κB p65 protein expression was significantly higher(P<0.01),hippocampal Caspase-1,NF-κB p65 protein expression was higher(P<0.05),in the agonist group compared with the medium dose group of Wenxinfang.Conclusions:1.High-fat diet feeding combined with chronic unpredictable mild stress allows the construction of an ApoE-/-mouse model of coronary heart disease with depression;2.Pathogenesis of coronary depression is closely related to NLRP3 inflammatory vesicles mediating inflammatory responses in the aorta and hippocampus;3.WenYang-Yixin therapy improved the degree of aortic atherosclerosis and depressive behavior in mice of coronary heart disease with depression;4.WenYang-Yixin therapy inhibited inflammatory responses in the aorta and hippocampus by a mechanism related to the inhibition of NLRP3 inflammatory vesicle activation and the reduction of NLRP3,ASC,Caspase-1,NF-κB protein expression and mRNA expression levels in the aorta and hippocampus.