The Effect Of Anthocyanins From Purple Sweet Potato Tuber On Cardiotoxicity Induced By Doxorubicin

Anthocyanin,a natural pigment,can protect or induce endogenous antioxidant defenses and have many health-care functions to the human body.Doxorubicin(DOX)is one of the effective drugs for cancer treatment.However,continuous administration of DOX can easily cause cardiomyocyte damage and myocardial fibrosis.Therefore,it is greatly significant to treat and regulate DOX-induced cardiotoxicity study via the use of plant-derived natural substances and antioxidants.In this study,anthocyanins were extracted from purple potato tuber by ethanol solvent extraction combined with ultrasonic assisted method,purified by macroporous resin AB-8 and Sephadex LH-20 column,anthocyanins from purple sweet potato tuber was identified by high-speed liquid chromatography-mass spectrometry(HPLC-DAD-ESI-MS).The protective effect of purple sweet potato tuber anthocyanins(PSPA)on DOX-induced H9C2 cardiomyocyte injury was studied by using DOX-induced H9C2 cardiomyocyte injury model.The influence mechanism of PSPA on DOX-induced cardiotoxicity of mice was studied by analyzing the secretion of injury markers in serum and heart tissue,and the changes of heart tissue morphology etc,which could provide a basis for the development and utilization of PSPA resources.(1)Isolation,purification and component identification of PSPA.After the PSPA were separated and purified by macroporous resin AB-8 and Sephadex LH-20 chromatographic columns,the content of PSPA was 6300 mg/100 g,which was higher than the crude extract.The component analysis results showed that it had 10 components,of which cyanidin and peonidin were the main components of PSPA,accounting for 62.9%and 21.46%of the total PSPA,followed by cyanidin-3-O-glucoside(6.5%)and cyanidin-3-O-sophoroside(2.8%),and a small amount of cyanidin-3-O-sophoroside,cyanidin-3-O-Malonyl hexosides,paeoniflorin derivatives,peonidin-3,5-O-diglucoside,peonidin-3-sophoroside-5-glucoside and caffeic acid 3-glucoside.(2)The effect of PSPA on the damage of H9C2 cardiomyocytes induced by DOX was studied.Cytotoxicity experiments showed that PSPA was not toxic to H9C2 cardiomyocytes in the range of 100-800 μg/mL.Compared with the normal control group(CON),the NO level in the DOX model group was significantly increased(p<0.05).The result indicated that the DOX model was successful,which induced H9C2 cardiomyocytes to release a large amount of inflammatory factor NO.The secretion of NO,TNF-α and trimethylamine-N-oxide(TMAO)in H9C2 cardiomyocytes of PSPA gavage group(DOX+PSPA)was dose-dependentl and significantly lower than that of DOX model group(p<0.05);and the activities of lactate dehydrogenase(LDH)and creatine kinase(CK)were significantly reduced in a dose-dependent manner(p<0.05).The results showed that PSPA could induce DOX by inhibiting the excessive release of DOX-induced inflammatory factors(NO and TNF-α)、TMAO、LDH and CK,and had protective effect on DOX-induced H9C2 cardiomyocyte damage.(3)The effect of PSPA on DOX-induced cardiotoxicity in mice was studied.The results showed that the body weight and the spleen index of the mice in the DOX model group were significantly decreased(p<0.05)and the heart index was significantly increased(p<0.05).However,the group of DOX+P SPA alleviated this phenomenon to a certain extent,and the group of high-dose PSPA(DOX+PSPA200)had the most significant effect(p<0.05).In the PSPA gavage group(DOX+PSPA),the amount of NO and MDA in the heart tissue,TNF-αand TMAO in the heart tissue and serum of the mice was significantly reduced in a dose-dependent manner(p<0.05);the activities of LDH and CK in heart tissue and serum were significantly reduced in a dose-dependent manner(p<0.05).The analysis of pathological sections of the heart tissue showed that myocardial rupture and vacuolar degeneration,a large number of inflammatory cell infiltration and myocardial cell granule degeneration in the DOX model group mice,showing obvious cardiac tissue damage morphology.However,in the group(DOX+PSPA),myocardial hypertrophy and interstitial hemorrhage were improved to varying degrees,which significantly reduced the changes in cardiac tissue caused by DOX.The results showed that DOX caused cardiotoxicity and damage in mice,and PSPA had protective and ameliorating effects on DOX-induced cardiotoxicity and damage in mice.Our results have proved that PSPA can effectively reduce and protect DOX-induced cardiotoxicity and myocardial injury,which could provide a certain experimental basis for PSPA as a functional food intervention on DOX-induced cardiotoxicity.