The Regulation Of Anthocyanins In Cr(Ⅵ)-induced Mitophagy In DF-1 Cells

Hexavalent chromium Cr(Ⅵ) is a very serious pollutant in environmental heavy metal pollution.The main reason is that the discharge of industrial residues pollutes the soil,natural environment and water sources.Cr(Ⅵ)enters the biological body,easily penetrates the outer cell membrane,enters the cell and exerts a strong oxidation effect,affects the normal life metabolism of the cell,and poses a huge threat to the body.A large number of data studies have shown that too much Cr(Ⅵ) can cause autophagy and lead to animal liver poisoning and kidney poisoning.Anthocyanins,also known as anthocyanins,have a variety of physiological effects such as anti-oxidation,improving liver function,preventing cardiovascular disease,anti-cancer,anti-inflammatory and protecting eyesight.Anthocyanins have a strong antioxidant capacity,mainly having a strong scavenging capacity for free radicals.In order to explore the regulatory effect of Lycium Ruthenicum Murr anthocyanins in Cr(Ⅵ)-induced mitochondrial autophagy in DF-1 cells,this experimental design was divided into control group,Cr(Ⅵ)group,Cr(Ⅵ)and different concentrations of Lycium Ruthenicum Murr anthocyanins the co-treatment group were used to detect the regulatory effect of anthocyanins in Cr(Ⅵ)exposure.Firstly,by detecting the cell survival rate to detect the degree of Cr(Ⅵ)-induced DF-1 cell damage and the protective effect of Lycium Ruthenicum Murr anthocyanins in the case of Cr(Ⅵ)-induced DF-1 cell damage.Secondly,by immunofluorescence and flow cytometry were used to detect reactive oxygen species and mitochondrial membrane potential to understand the degree of anthocyanin-induced mitochondrial damage in DF-1 cells in Cr(Ⅵ).Detect autophagy-related proteins(p62,LC3,ATG5,Beclin-1),mitophagy-related protein(PINK1)and the recruitment of Parkin on the mitochondria by protein immunoblotting and ELISA kits,and explore the role of anthocyanins in Cr(Ⅵ)induces autophagy in DF-1 cells and the role of mitophagy.The test results found that the addition of anthocyanins can alleviate mitochondrial damage caused by Cr(Ⅵ),induce cellular oxidative stress and mitochondrial depolarization.Cr(Ⅵ)causes the overexpression of autophagy proteins(LC3,Beclin-1,Atg5)and reduces the expression of autophagy protein p62.Anthocyanins can also inhibit the transfer of Parkin from cytoplasm to mitochondria,thereby reducing the recruitment of Parkin on mitochondria caused by hexavalent chromium.In short,the above test results show that Cr(Ⅵ) entering DF-1 cells will cause a certain degree of damage to the mitochondria in DF-1 cells,cause mitophagy and activate the PINK1/Parkin pathway.When Lycium Ruthenicum Murr anthocyanins are added,Cr(Ⅵ)-induced mitochondrial damage and mitophagy in DF-1 cells can be relieved,and Parkin’s transfer from cytoplasm to mitochondria and the recruitment of PINK1 on mitochondria can be reduced.