The Mechanism Of Sodium Creatine Phosphate Alleviate Doxorubicin Cardiotoxicity

OBJECTIVE:Discuss the protective effect of sodium creatine ph-osphate on myocardial injury induced by doxorubicin and explain the core reg-ulation of tak1 in it.METHODS:?1?In vivo experiment,male SD rats were randomly divided into model group?DOX group?,drug treatment group?CP+D OX group?and normal control group?CON group?.Rats were ip injected with DOX 2 mg�kg^-1once every other day,for 7 times severed as DOX group,were ip injected with CP 200 mg�kg^-11 once every other day for 3 times,then ip in-j ected with DOX 2 mg�kg^-11 plus CP 200 mg�kg^-11 for 7 times severed as CP+D OX group and normal control group were ip injected with normal saline 5 m L�kg^-11 for times as same as CP+DOX group.?2?In vitro experiment,DOX 1?mol�L^-11 treated alone or jointly with CP 1 mmol�L^-1?n-acetyl-l-cysteine?NAC?0.5 mmol�L^-1,TAK1 inhibitor 5z-7-oxozeaenol to treat cardiomyocytes 24 h,E-lectrocardiogram?ECG?was used to observe the cardiac function of rats,and HE staining was used to observe the pathological changes of myocardium.The accumulation of collagen fibers in rat myocardial tissue was detected by Mass-on staining.The apoptosis level of H9C2 cardiomyocytes were detected JC-1mitochondrial membrane potential method and the activity H9C2 cardiomyocyt-es were detected by CCK8 kit.The expression of antioxidant enzyme superoxi-de dismutase?SOD2?,nuclear factor e related factor 2?Nrf2?,forkhead protein Foxo3a and TGF-?activated kinase 1?TAK1?,Ca²⁺mediation kinase?CaMKII?,receptor interacting protein 3?RIP3?,apoptosis-related protein b lymphocyte tu-mor-2 gene-related protein?Bax?,lymphoma-2 gene?Bcl2?,cleaved-caspase3 in myocardial tissue and cardiomyocyte of rats were detected by western blot.RESULTS:?1?In vivo experiment,The results of echocardiography showed t-hat DOX group had significantly lower ejection fraction,shortening fraction an-d cardiac output than the blank control group?P<0.05?,The ejection fraction s hortening fraction and cardiac output of rats in CP+DOX group compared with DOX group?P<0.05?;The results of HE,Masson staining showed that:When compared with the normal control group,the arrangement of muscle fibers in DOX group was disordered,the tissue cells had obvious vacuolar-like changes,the accumulation of collagen fibers increased,and the apoptosis increased sig-nificantly?P<0.05?.The muscle fibers in CP+DOX group were arranged neatly,vacuoles disappeared,cell structure was clear,collagen fibers accumulation dec-reased?P<0.05?;Western blot results showed that compared with normal contr-ol group,In DOX group,The expression of oxidative stress-related protein Nrf2?SOD?Foxo3a decreased significantly and TAK1 expression decreased?P<0.05?.The expression of necrosis-related protein RIP3?mitochondrial damage-relat ed protein CaMKII increased significantly?P<0.05?.Up-regulated expression of apoptosis-related protein cleaved-caspase3?Bax and down-regulat-ed expression of anti-apoptotic protein Bcl2?P<0.05?.?2?In vitro experiment,CCK8 cell vi-ability test results showed that sodium creatine phosphate can significantly imp-rove cardiomyocyte viability.The results merge JC-1 mitochondrial membrane potential experiments shown that,compared with the normal control group,DOX group had more green fluorescence,decreased mitochondrial membrane potenti-al and cell apoptosis clearly.When compared with DOX group,green fluoresc-ence decreased,red fluorescence increased,mitochondrial membrane potential i-ncreased and apoptosis decreased in CP+DOX group,H9C2 results of cardiom-yocyte experiment were consistent with those of tissues.The results of Western blotting showed that the Myocytes treated with different concentrations DOX+CP showed that the expression of TAK1 increased with the increase of TAK1concentration and decreased with the increase of DOX concentration.Expression of Nrf2,SOD,TAK1 and Bcl2 in DOX group was down-regulated compared with cell control group.The expression of RIP3,CaMKII,cleaved-caspase3 and Bax was up-regulated.In NAC group,the expression of Nrf2,SOD,Foxo3a and TAK1 was up-regulated,and the expression of RIP3,CaMKII,and cleaved-caspase3 were down-regulated,the histone expression of TAK1 inhibitor 5z-7-ox was consistent with that of the control group,The expression of Nrf2,SOD,Foxo3a,TAK1 and Bcl2 were down-regulated in the 5z-7-ox+DOX group co-mpared with the DOX group,RIP3,CaMKII,cleaved-caspase3 and Bax expre-ssion were significantly up-regulated.Conclusion:Sodium creatine phosphate not only improves cardiac dysfunction induced by doxorubicin,also reduce the antioxidant capacity that DOX induce myocardial tissue and cardiomyocytes,and promote TAK1 expression.Improving DOX induced cardiomyocytes apoptosis.