The Role Of Hydrogen Sulfide In The Model Of Doxorubicine-induced Injury Of H9C2 Cell Line

BackgroundDoxorubicine is a widely-used anti-tumor drugs especially in the treatment of breast cancer and in the field of hemotology oncology.However,its life-threatening cariac complication definitely limits its clinical use.it has been demonstrated that the mitochondria is the target of Doxorubicine,which is regarded as the main mechanism of cardiac toxicity induced by Doxorubicine.Hydrogen Sulfide is the third gas transmitter since CO,NO has been discovered and has been investigated to take part in many essential physiological or pathological processes in the body, such as ischemia-reperfusion injury vessel-dilation,anti-inflammation.In addition, some research groups have found that Hydrogen Sulfide can protect the heart from ischemia-reperfusion injury through protection of mitochondria of cardiomyocytes.Objectivewe try to explore whether H2S can attenuate the cardiac toxicity induced by Doxorubicine through mitochondrial pathway.MethodsH9C2 cells (rat cardiomyoblast cells) incubated with DOX and NaHS (H2S donor) were observed under inverted microscope and analyzed for cell viability. Secondly, the mitochondrial potential of cardiomyocytes was examined by Fluorescence Microscopy and Flow cytometry using Rh123 and JC-1 staining respectively.In addition, we used Western-blot to detect the expression of Akt protein.Results1. Doxorubicine can dose-dependently reduced cell viability and Hydrogen Sulfide had no obvious effects on cardiomyocytes viability up to 200μmol/L level but can alleviate the cell viability when it was co-cultured with Dox at proper concentration.2. Hydrogen Sulfide can effectively reverse the collapse of mitochondrial potential induced by DOX at the concentration of 100μmol/L.Akt expression which was decreased by DOX was enhanced by H2S.ConclusionAt appropriate concentration and time,Hydrogen sulfide can reverse the declining of mitochondrial function demonstrated by cell viability and mitochondrial potential induced by Doxorubicine. In addition, H2S can enhance the expression of total Akt Protein. Consequently ,we conservatively inferred that the PI3K-Akt pathway was involved in the process.