Royal Acid Relieves Myocardial Autophagy Damage Caused By Doxorubicin And Promotes Tumor Cell Apoptosis

Doxorubicin(Dox)as a kind of classic anthracycline-based drugs,has been used in clinical treatment of tumor,but its side effects will cause certain damage to the heart.The exact mechanism of cardiac toxicity induced by Dox remains unclear,but recent studies suggest that autophagy and apoptosis are involved in the cardiac toxicity.In order to reduce the cardiac toxicity in clinical practice,the search for Chinese herbal medicine antagonizing cardiac toxicity of Dox is imminent except reducing the use of Dox.Royal jelly acid(10-Hydroxy-2-decenoicacid,10-HDA)is a kind of unsaturated fatty acid with high stability isolated from the royal jelly,which was proved to have antibiotic,sterilization,antiinflammatory,antitumor,antiradiation,antiulcer,enhance immunity,inhibit fat synthesis,decreasing blood lipids and other activities.At present,there is not a whole lot of research into 10-HDA used to tumor treatment and we have not seen the reports about if 10-HDA has protective effect on myocardial cell injury induced by Dox.In this study,we use H9c2 cells and Hela cells as the object to observe the protective effect of 10-HDA on myocardial injury induced by Dox on cell level,and to explore its possible molecular mechanisms.The main results were as follows.Treated H9c2 cells with pure ethanol(solvent),2,4,8,12 mM 10-HDA for 12 hours,and then observed that there was no cellular morphology difference in invertion flurescence microscope in addition to the concentration of 12 mM which led to a large number of cell death.Then we treated H9c2 cell respectively With different concentrations of 10-HDA such as 0,0.1,0.2,0.5,1,2,4,8 mM at the same time,the results of western blot show that 10-HDA can reduce autophagy of H9c2 cell and high concentrations of 10-HDA can reduce cell apoptosis.We adopted 1,2,4,8 mM as 10-HDA dosing concentration gradient in the following treatment experiments and preincubation experiments.The results of treatment experiments show that 10-HDA can relieve the autophagy of H9c2 cells caused by Dox,but may not be able to alleviate the apoptosis,even can increase the apoptosis after 24 hours.At the same time,CCK-8 dynamic testing found 10-HDA is hard to enhance its vigor after firstly treating cells with Dox but it also did not reduce its vitality any more.Under the condition of preincubation,10-HDA can also reduce autophagy of H9c2 cell but may not be able to alleviate the apoptosis,but also will not increase.At the same time,found that preincubation with 1-8 mM 10-HDA can reduce the cytotoxicity caused by Dox and increase cell vitality.We firstly groped for Dox concentration gradient in Hela cell and chose 1 pM as drug concentration in the following experiments,because the autophagy would be at the highest degree and apoptosis and cell vitality would be in normal level.By 10-HDA concentration gradient experiment,we found that the cell vitality almost unaffected under the condition of concentration 1,2,4,8 mM,so we continue to choose this concentration gradient in the subsequent treatment and preincubation experiments.Treatment experiments and preincubation expwufueriments show that 10-HDA has a dose-dependent effect on the treatment of myocardial injury caused by Dox in Hela cell.Autophagy decreased and apoptosis increased with the increase of dosage.After that,we chose 5-Fluorouracil(5-FU)and cisplatin(cis-Dichlorodiamineplatinum,DDP)to do further research and found that 10-HDA also can relieve autophagy and promote apoptosis in combination with other medicines in Hela cell.This means 10-HDA can be used as a broad-spectrum ancillary in tumor therapy.