The Protective Effect Of Anthocyanin In Apoptosis Of DF-1 Induced By Arsanilic Acid

Arsenic is a common metal-like toxic element which is often used as a herbicide and insecticide.Arsanilic acid is a kind of organic arsenic synthesized by the reaction of aniline and arsenic acid,which is less toxic than inorganic arsenic,and is commonly used as a feed additive for livestock and poultry.However,improper use of arsanili acid as a feed additive can also cause chronic poisoning in animal organism.Anthocyanin is a kind of flavonoids extracted from the mature fruit of plants,which has the effect of antioxidation,free radical scavenging and anti-inflammatory.In order to study the toxicity of arsanilic acid to DF-1 cells and the protective effect of anthocyanins as antioxidant on apoptosis of DF-1 cells induced by p-aminobenzene arsenic acid.This paper construsted a toxic model of chicken embryo fibroblast cell line DF-1 in our experiment.The model was treated with arsanilic acid and anthocyanin for 24 hours.Firstly,the concentration of arsanilic acid and the concentration of anthocyanin protected cells were determined by cell viability,and the cell apoptosis rate was detected by flow cytometry.The changes of intracellular reactive oxygen species and mitochondrial membrane potential were used to verify that arsanilic acid could induce apoptosis and the protective effect of anthocyanins on cells.Detection of MDA by colorimetry to explain the changes of intracellular oxidative stress.Laser confocal technique was employed to detected the morphological changes of apoptosis.Moreover,western blotting technique was used to analyze the expression of caspase-3 protein.The results showed that 10 mM arsanilic acid had obvious toxic effect on cells and induced apoptosis.50 μ g/mL and 100 μ g/m L anthocyanins could significantly antagonize the phenomenon such as damage of cells,increase of apoptosis rate,production of reactive oxygen species and MDA in cells induced by arsanilic acid.Moreover,anthocyanin can be effectively inhibited the decrease of mitochondrial membrane potential and activation of caspase-3.In summary,arsanilic acid could induced apoptosis of DF-1 cells,and anthocyanins could reversed arsanilic acid-induced apoptosis by restoring mitochondrial membrane potential,reducing the production of ROS and MDA in cells and caspase-3 and other related apoptosis factors.