Aconitine Induces Apoptosis In H9c2 Cardiac Cells Via Mitochondria-mediated Pathway

Objective: Aconitine,a diterpenoid alkaloids derived from Aconitum plants,is widely employed to treat various diseases.Aconitine poisoning accidents have occurred occasionally caused by inadequate consumption of aconitine in recent years.Moreover,the cardiotoxicity and neurotoxicity of aconitine have been reported in the literature.In order to further clarify the toxic effects of aconitine and provide relevant theoretical basis for aconitine poisoning cases in forensic practice,H9c2 cardiac cells were used as a model in our present study,to explore the toxic effects of aconitine and relevant mechanism of cell apoptosis induced by mitochondrial pathway.Methods: H9c2 cardiac cells were used as a model in vitro exposed to aconitine.H9c2 cell apoptosis induced by aconitine was detected by a Cell Counting kit-8 assay,DAPI staining,Annexin V-FITC/propidiumiodide double staining and western blotting.The effects of aconitine on reactive oxygen species levels and mitochondrial membrane potential were confirmed by fluorescence microscopy and flow cytometry.In addition,ATP contents were determined using a ATP-dependent bioluminescence assay kit.The levels of peroxisome proliferator activated receptor ? co-activator 1?(PGC-1?)expression and apoptosis-associated proteins including Caspase-3,Bcl-2,Bax and Cytochrome c were also assessed.Results: Aconitine could inhibit cell viability,induce mitochondrial dysfunctions including increase ROS generation,decrease ATP contents and ?? m,decrease PGC-1? expression,upregulate Cytochrome c,Bax and Caspase-3,and downregulate Bcl-2,suggesting that aconitine could induce apoptosis through mitochondria-mediated pathway in H9c2 cardiac cells.Conclusion: Aconitine could inhibit cell viability,cause nuclear morphological changes,induce cell apoptosis,suggesting that aconitine has obvious toxic effects on cardiac cells.Moreover,aconitine could increase ROS generation,decrease ATP contents,?? m and PGC-1? expression,indicating aconitine has certain damage to mitochondria.Aconitine upregulated Cytochrome c,Bax and Caspase-3,and downregulated Bcl-2,showing that aconitine could induce cell apoptosis.In summary,aconitine could induce apoptosis of H9c2 cardiac cells at least in part via mitochondria-dependent apoptotic pathway.