| Background:Heart failure?HF?is the end stage of various cardiovascular diseases,such as hypertension,coronary heart disease and so on.The incidence of HF is increasing year by year.It is a problem that needs to be solved in cardiovascular field and has become a public health problem.Its pathogenesis has penetrated into many fields,such as neuroendocrine system,humoral regulatory factors and signal transduction,but it has not been fully elucidated.With the deepening of research,it has been found that heart failure is accompanied by the activation of inflammatory in recent years.Therefore,it can be seen that inflammatory plays an extremely important role in the pathophysiological process of heart failure.Inflammation deteriorates cardiac function,and the deterioration of cardiac function further aggravates the inflamma-tion,forming a vicious circle between the two.Levosimendan is a new positive inotropic drug.It is a calcium sensitizer and combines with troponin to increase myocardial contractility.Avgeropoulou et al.confirmed that levosimendan was not only effective in preventing heart failure,but also significantly reduced the level of inflammatory factors compared with dopamine.Feola et al.showed that compared with furosemide,levosimendan not only improved the symptoms of heart failure,but also significantly reduced the serum inflammatory factors in patients with heart failure.After the intervention of levosimendan,the level of inflammatory factors decreased significantly.It can be seen that levosimendan has anti-inflammatory effect,but its specific mechanism is not clear.NF-KB proteins are heterodimers composed of two Rel family proteins.They are multipotent transcription factors.They contain DNA binding regions at the amino acid residues.They interact with DNA binding,NF-KB inhibitor proteins and carry nuclear localization signals.Activated KB sites at the promoter sites of NF-KB binding activate transcription expression.IKB protein consists of three parts:N-terminal region,C-terminal region and internal region.When IKB protein is phosphorylated,the trimer complex of NF-KB protein and IKB protein decomposesand releases the NF-KB protein,so that it can transfer from cytoplasm to nucleus.Nuclear factor-kappa B?NF-KB?signaling pathway exists widely in the body.It is composed of the protein of the NF-KB family and its inhibitory protein IKB?NF-kB?family.It plays an important regulatory role in various inflammatory and immune processes.TLR4 receptor is a transmembrane receptor protein,which consists of three regions.When TLR4 receptor binds to the corresponding ligands,it forms receptor ligand structure to further activate the NF-KB signaling pathway,and then affect the expression of inflammatory cytokines.Gene expression of inflammatory factors such as IL-1,IL-6 and TNF-alpha.TLR4/NF-KB signaling pathway is an important inflammatory pathway.It was found that levosimendan can prevent heart failure and reduce serum inflammatory factors.In this study,we first selected patients with heart failure to give levosimendan intervention to analyze the changes of inflammatory markers in blood;secondly,levosimendan intervention to heart failure rats to further detect the changes of inflammatory markers in blood,the expression of IL-6 and TLR4 in rat myocardium,the expression of IKB,NF-KB P65 and calmodulin protein.To explore the anti-inflammatory mechanism of levosimendan will provide a theoretical basis for elucidating the anti-inflammatory mechanism of levosimendan.Part?: Effect of levosimendan on early efficacy and inflammatory in patients with heart failure Objective:The patients with heart failure with lower ejection fraction?LVEF?40%?NHYA III-IV were selected as the research object,and the intervention of levosimendan was given to observe the early clinical effect of levosimendan on heart failure and the influence of serum inflammatory factors.Methods:From May 2016 to December 2018,220 patients with NHYA III-IV heart failure with ejection fraction less than 40% were randomly divided into two groups:Levosimendan group?110 cases,of which 10 cases were missing?and control group?110 cases,of which 10 cases were missing?.The control group was treated with diuretic,cardiotonic,vasodilator and other anti-heart failure therapies;the levosimendan group was treated with levosimendan 12.5 mg on the basis of the above anti-heart failure therapies?starting with a loading dose of 6 ug/kg for 10 minutes,then 0.05-0.2 ug/kg/min continuous intravenous pump for 24 hours?,and the dosage was adjusted according to the specific conditions of patients.Monitoring indicators:The changes of blood pressure,heart rate,BNP,IL-6,white blood cells,neutrophils,creatinine and LVEF were observed before and 24 hours after treatment.Results:1.General data: There was no significant difference in the basic data between the levosimendan group and the control group at admission?p > 0.05?.The SBP and DBP of control group and levosimendan group decreased after treatment,and the difference was statistically significant?p < 0.05?.The HR of control group and levosimendan group decreased after treatment,but there was no significant difference between the two groups?p > 0.05?.2.Early evaluation of clinical efficacy: BNP in control group and levosimendan group decreased after treatment,and there was no significant difference between control group and before treatment?p > 0.05?;BNP in levosimendan group decreased significantly compared with before treatment?p < 0.001?.The LVEF values of control group and levosimendan group increased after treatment,but there was no significant difference between the two groups?p > 0.05?.3.Inflammation index: IL-6 in control group and levosimendan group decreased after treatment,and IL-6 in levosimendan group decreased significantly compared with before treatment?p < 0.001?.Leukocyte in control group and levosimendan group decreased after treatment,and levosimendan group decreased significantly?p <0.001?.The neutrophils in the control group and the levosimendan group decreased after treatment,and the difference was statistically significant.The levosimendan group?p < 0.001?and the control group?p < 0.05?.4.Influences of incidence of atrial fibrillation: The incidence of atrial fibrillation in levosimendan group was higher than that in control group.The results of2 test showed that there was no significant difference between the two groups?>0.05?.5.Effect on creatinine: After treatment,creatinine increased in both control group and levosimendan group,and there was no significant difference in creatinine increase between the two groups?p > 0.05?.Conclusion:1.Levosimendan reduces IL-6,leukocytes and neutrophils in patients with heart failure.2.Levosimendan decreased BNP level in patients with heart failure;compared with the control group,left Simendan group increased LVEF,but there was no significant difference.3.The incidence of atrial fibrillation in levosimendan group was higher than that in control group,but there was no significant difference.4.Levosimendan has little effect on serum creatinine in patients with heart failure.Part?: Effect of levosimendan on inflammatory in rats with heart failureObjective:In this study,isoproterenol was used to establish heart failure model in rats.On this basis,levosimendan was used to intervene.The changes of inflammatory factors in blood,myocardial tissue and protein expression in rats with heart failure related to TLR4/NF-KB signaling pathway were observed from the "heart failure rat-serum inflammatory factor-myocardial tissue-signaling pathway" route,so as to explore the effect of levosimendan on heart failure.Inflammatory factors in rats with heart failure.Method:8-10 weeks SPF male SD rats?body mass 200-250g?were divided into two groups: experimental group rats were subcutaneously injected with isoproterenol 3mg/d for 4 consecutive weeks;control group rats were subcutaneously injected with saline 3 mg/d for 4 consecutive weeks.Four weeks later,the heart function of rats in the two groups was measured by transthoracic echocardiography using Vevo 2100 small animal color Doppler of Visual Sonics Company.The serum BNP of the two groups was detected by ELISA kit.The success of rat heart failure model was evaluated by color Doppler echocardiography and BNP.The rats with heart failure were divided into the following three groups: rats in the heart failure group were intraperitoneally injected with 5% glucose 2.0 mg/kg,rats in the levosimendan group were intraperitoneally injected with levosimendan 2.0 mg/kg,rats in the TLR4 receptor blocker + levosimendan group were intravenously injected with TLR4 receptor blocker 0.5 mg/kg + levosimendan 2.0 mg/kg,rats in the control group were intraperitoneally injected with 5% glucose 2.0 mg/kg,and rats in the second group Blood samples were taken from rat hearts directly to detect white blood cells and neutrophils;serum IL-6 and BNP were measured by ELISA;myocardial tissue was taken for detection by RT-PCR;expression of IL-6 and TLR4 in myocardial tissue was detected;expression of IKB protein and nuclear protein NF-KB P65 was detected by Western Blot method.Results:1.The ventricular enlargement and cardiac function of rats in isoproterenol group were decreased,and the serum BNP value was significantly higher.Combining with the results of echocardiography and BNP,heart failure of rats in isoproterenol group was successfully established.2.The results of BNP in TLR4 blocker + levosimendan group and levosimendan group were not abnormal.BNP in heart failure group was significantly higher than that in control group and levosimendan group?p < 0.001?,and BNP in levosimendan group was significantly higher than that in control group?p < 0.05?.3.The results of IL-6 in TLR4 blocker + levosimendan group and heart failure group were not abnormal.IL-6 in heart failure group was higher than that in control group and levosimendan group?p < 0.05?,while IL-6 in levosimendan group was higher than that in control group,but there was no significant difference?p > 0.05?.4.There were no abnormal results in TLR4 blocker + levosimendan group and heart failure group.Leukocyte count in heart failure group was significantly higher than that in control group?p < 0.001?.Leukocyte count in heart failure group was significantly higher than that in levosimendan group?p < 0.05?.There was no significant difference between levosimendan group and control group?p > 0.05?.There was no significant difference in neutrophils between the four groups.5.There was no abnormality in the results of NF-KB P65 protein and heart failure in TLR4 blocker + levosimendan group.The expression of NF-KB P65 protein in levosimendan group was lower than that in heart failure group?p < 0.05?,and the expression of NF-KB P65 protein in levosimendan group was higher than that in control group?p < 0.05?.The results of IKB protein in TLR4 blocker +levosimendan group and heart failure group were not abnormal.IKB protein in left Simendan group was lower than that in heart failure group?p < 0.05?;IKB protein in left Simendan group was higher than that in control group?p < 0.05?;IKB protein in heart failure group was higher than that in control group?p < 0.05?.6.There were no abnormalities in IL-6 gene expression and heart failure in TLR4 blocker + levosimendan group.The expression of IL-6 in levosimendan group was significantly lower than that in heart failure group?p < 0.05?;the expression of IL-6 in levosimendan group was higher than that in control group,but there was no significant difference between the two groups?p > 0.05?;the expression of IL-6 in heart failure group was significantly higher than that in control group?p < 0.05?.The expression of TLR4 in TLR4 blocker + levosimendan group and heart failure group were not abnormal.The expression of TLR4 in levosimendan group was lower than that in heart failure group,the difference between the two groups was statistically significant?p < 0.05?;the expression of TLR4 in levosimendan group was higher than that in control group?p < 0.05?;the expression of TLR4 in heart failure group was higher than that in control group,and the difference between the two groups was statistically significant?p < 0.05?.Conclusion:1.Levosimendan corrects isoproterenol-induced heart failure.2.Levosimendan inhibits the inflammatory factors of cardiac failure induced by isoproterenol.3.Levosimendan inhibits the inflammation of cardiac failure induced by isoproterenol through TLR4/NF-KB signaling pathway. |
PDF Full Text Request