| Background:Cytochrome P450 2E1(CYP2E1)is a heme protein encoded by the superfamily of cytochrome p-450 gene.CYP2E1 is up-regulated in a variety of heart diseases,causing damage mainly through reactive oxygen species.Studies have shown that in mice,increased expression of CYP2E1 can induce myocardial cell apoptosis,while decreased expression of CYP2E1 can alleviate the pathological development of dilated cardiomyopathy.However,the role and mechanism of targeted inhibition of CYP2E1 in the treatment of dilated cardiomyopathy remains unclear.This study was to research the diene propyl disulfide(Diallyl sulfide,DAS)targeted inhibiting CYP2E1 on treatment effect and possible mechanism of DCM.Methods:(1)Animal grouping and drug administration:the 4-month-old cTnTR141W transgenic mice and the negative wild type mice were randomly divided into 6 groups.The 6 groups were:Non-transgenic littermates(NTG),a group of cTnTR141W transgenic mice,DAS low-dose group(200 mg/kg),DAS high-dose group(400 mg/kg)and positive drug control group(Enalaprilat,0.76 mg/kg).(2)Phenotypic analysis of the morphology and function of mouse heart at the global,microscopic and ultrastructural levels:ultrasonic image analysis was used to observe the overall function and structure of mouse heart.H&E staining,Masson staining and transmission electron microscopy were used to analyze the myocardial morphology.The expression of myocardial fibrosis markers in mice was analyzed by polymerase chain reaction.(3)Biochemical and molecular biological techniques were used to analyze the molecular mechanism of DAS involvement in oxidative stress and apoptosis in myocardium in mice with heart expansion disease:biochemical analysis of mouse ROS markers,namely glutathione,hydrogen peroxide,and propionic acid Dialdehyde was measured.Molecular biological techniques:Tunel detection and Western Blot were used to determine myocardial cell apoptosis and related apoptotic marker molecules,including Procaspase-9,Activated caspase 9,Cytochrome c,Procaspase 3 and Activated caspase 3.Results(1)On the overall level,ultrasound image analysis showed that the pathological phenotypes of cTnTR141W dilated cardiomyopathy model mice,including ventricular cavity dilation,ventricular wall thinning,and ventricular function weakening,were significantly improved after DAS treatment.On the microscopic level,by H&E and Masson histological staining results showed that the DAS processing,cTnTR141W DCM myocardial micro array model mice improved,myocardial cell interstitial collagen deposition improved,at the same time,the mRNA level detection of collagen type III has carried on the proof to the improvement of the collagen deposition.At the ultrastructural level,transmission electron microscopy showed that after DAS treatment,the myocardial ultrastructure damage of cTnTR141w dilated cardiomyopathy model mice was significantly improved,including myocardial fiber fracture and dissolution,myoplasmic reticulum expansion,mitochondrial swelling,cristae loss and vacuolization.At the same time,after enalaprilat treatment,the cardiac function and morphology,collagen deposition in the intercellular space of cardiomyopathy model mice of cTnTR141W significantly improved.(2)Through the detection of oxidative stress and key molecules of apoptotic signals,it was shown that after DAS treatment,the levels of hydrogen peroxide and malondialdehyde were decreased,the levels of reduced glutathione were significantly increased,and the levels of oxidative stress were significantly improved in the myocardial tissue of cTnTR141W dilated cardiomyopathy model mice.At the same time,the apoptosis number of cardiomyocytes in cTnTR141W dilated cardiomyopathy model mice was significantly inhibited after DAS treatment,and the activation of Caspases 9 and 3 in cardiac tissue was significantly inhibited,and the release of Cytochrome c was also significantly improved.(3)Although enalapril had no significant effect on the level of myocardial oxidative stress in cTnTR141W DCM mice,it could reduce the number of myocardial cell apoptosis in cTnTR141W DCM mice by inhibiting the mitochondrial apoptosis pathway.Moreover,enalaprilat treatment significantly improved the structure,function and pathological changes of the heart,especially the changes of the microstructure and ultrastructure.Conclusion:DAS treatment can significantly improve the pathological phenotype of cardiomyopathy in cTnTR141W dilated cardiomyopathy model mice,including ventricular dilatation,ventricular wall thinning,myocardial fibrosis and cardiac function weakening.DAS after processing by competitive inhibition CYP2E1 and reduce the myocardial oxidative stress level,and inhibition of mitochondrial apoptosis pathway,which reduces the myocardial cells of cytochrome c from mitochondria to the amount of release in the cytoplasm,and inhibition of cytochrome c in the cytoplasm and the apoptotic protease activating factor 1,dATP and Procaspase-9 combination,thus inhibiting Caspase 9 and Caspase 3 activation,and inhibit the apoptosis of myocardial cells.Inhibition of CYP2E1 may be a valuable therapeutic strategy to control the occurrence and development of cardiovascular diseases involving up-regulation of CYP2E1.In addition,the development of DAS analogue with strong inhibition of CYP2E1 and high bioavailability can provide clues and research directions for the prevention and treatment of dilated cardiomyopathy and heart failureBackground: Alzheimer’s disease is a complex neurodegenerative disease.Due to the complexity of its molecular pathogenesis and the interaction of various factors,the etiology and pathogenesis of AD have not been fully elucidated.Therefore,treatments for AD needs to be developed.Evodiamine extracted from the Chinese herbal medicine Evodia is a quinolone alkaloid.Studies have found that Evodiamine can inhibit inflammation and excessive activation of microglia in the brain,improve the learning and transgenic mice of APPswe/PSlAE9 Spatial memory ability.Methods: Evodiamine and its derivatives were synthesized by condensation reaction mediated by EDCI.In vitro,the cytotoxicity of evodiamine derivatives to SH-SY5 Y and HepG2 was detected by cell proliferation detection,and the neuroprotective effect of evodiamine derivatives on the damage of SH-SY5 Y cells induced by hydrogen peroxide was further detected.In vivo,and through the APPswe/PSlAE9 AD model mice Morris water maze experiment testing evodiamine to model the effect of learning and spatial memory ability in mice.Results: Evodiamine derivatives containing 1 thiogen and 21 different substituents were synthesized effectively by ECDI-mediated condensation reaction.Some low cytotoxicity of evodiamine derivatives,has obvious nerve protective effect,can significantly improve APPswe/PSlAE9 mice learning and spatial memory ability.Conclusion: A series of new and more effective derivatives of evodiamine have been synthesized effectively,and the therapeutic effect of evodiamine derivatives on AD has been further discussed,which provides an idea for the development of new and effective evodiamine derivatives in the treatment of AD. |
