Effect Of FRRT On Autophagy And Apoptosis Of Myocardial Cells Induced By Doxorubicin

BackgroundDoxorubicin(DOX)is used in the treatment of clinical common malignant tumors.The efficacy of DOX is remarable,however,the cardiac toxicity reduce the quality of life of patients and limited it's clinic application.There are several signaling pathways that induce the cardiotoxicity by DOX.Including the autophagy and apoptosis of cells,those can cause to cardiotoxicity.The pear fruit contains a lot of pharmacological components,such as flavonoids.The total flavonoids content is 6.8 g/kg,which is up to8.0g/kg.At present,researches have indicated that flavonoids has play an important role of dilating coronary artery and protecting cardiovascular.However,people know very little about the flavonoid's protective efficacy on cardiotoxicity caused by DOX.In this study,we aim to investigate the efficacy and the mechanism of FRRT in the progress of apoptosis and autophagy in doxorubicin-induced cardiotoxicity by cultured myocardial cells.ObjectiveTo investigate the effect and mechanism of FRRT on autophagy and apoptosis in doxorubicin-induced cardiotoxicity.MethodsMyocardial cells were separated and cultured from newborn 1-3 days of rat and H9C2 myocardial cell line.The cardiotoxicity model were established with 5?mol/L doxorubicin incubation,and FRRT group were intervened with FRRT incubation in advance.Ultrastructure of H9C2 Myocardia L cells were observed with transmission electron microscope.The distribution and expressions of autophagy related genes(LC3-II?P62)and apoptosis related genes(Bax?P53 and Bcl-2)in myocardial cells weredetected with Western Blotting and Immunofluorescence,and the results of Western Blotting were analyzed semi-quantitatively used by Image J and Grapf Pad Prism 5.Results1.Morphological observation1.1 Cell growth is poor status in DOX group,most cells shrink and present high refractive circles.The cells grew well and most of them showed long fusiform shape in control group,and they can regular pulsation.Only part of the cells shrink and present high refractive circles,most of them are still long fusiform shape.1.2 The ultrastructure of myocardial cells showed a large number of autophagy and lipofuscin,structure of various organelles is ambiguous,and were of poor condition in DOX group,and no lipofuscin and a small amount of autophagy in FRRT group,various organelle structures are clearly visible.2.The results of Immunofluorescence2.1 Autophagy associated protein detection revealed,LC3-II was mainly expressed in the DOX group,and the expression of LC3-II in the Control group was not obvious,and the FRRT group was expressed in the cytoplasm.P62 was very few expressed cytoplasm in the DOX group.In the Control group,it was expressed clearly in the cytoplasm of most cardiomyocytes,and some cells expressed P62 in the FRRT group.There was no positive expression of LC3-II and P62 in negative control group.2.2 Apoptosis related protein detection revealed,Bax was evenly expressed in the cytoplasm in the DOX group,and no obvious expression was detected in the Control group and FRRT group.P53 in the DOX group showed a large number of expression in the cell nucleus,and the expression in the Control group was not obvious,and the FRRT group showed a small amount of expression.Bcl-2 was expressed in small amounts in the surrounding cytoplasm in the DOX group.The Control group and the FRRT group expressed clearly in the cytoplasm around the cell nucleus.There was no positive expression of Bax?P53 and Bcl-2 in negative control group.3.The results of Western Blotting3.1 Autophagy associated protein detection revealed,LC3-II expressed in DOX group than the Control group and the FRRT group increased,the comparison between groups,statistically significant difference(** P < 0.01),FRRT group than the Control group expression was slightly increased,but the comparison between groups,no statistically significant difference(P > 0.05).P62 expressed in DOX group than the Control group and the FRRT group reduced,the comparison between groups,statistically significant difference(* *P < 0.01),FRRT group than the Control group expression was slightly reduced,but the comparison between groups,no statistically significant difference(P >0.05).3.2 Apoptosis related proteins tests revealed,Bax expressed in DOX group than the Control group and the FRRT group increased,the comparison between groups,statistically significant difference(* P < 0.05),FRRT group than the Control group expression was slightly increased,but the comparison between groups,no statistically significant difference(P > 0.05).P53 expressed in DOX group than the Control group and the FRRT group increased,compared with the Control group,the expression of FRRT group was not significantly increased,and the difference was not statistically significant(P>0.05).Bcl-2expressed in DOX group than the Control group and the FRRT group reduced,the comparison between groups,statistically significant difference(** P < 0.01),FRRT group than the Control group expression was reduced,the comparison between groups,statistically significant difference(* P < 0.05).ConclusionsFRRT'effect from doxorubicin-induced cardiotoxicity by reducing the autophagy nd apoptosis in myocardial cells.