The Role Of Drp-1 In The Damage Of Primary Chicken Hepatocytes Injury Induced By High Copper

Copper is one of the important elements to organism.It participates in some important physiological processes,such as cellular respiration process,mutual transmission of neurotransmitters,anti-oxidative stress reaction and iron absorption.Previous studies have shown that high concentrations of copper can seriously affect the mitochondrial respiratory function,by inducing the occurrence of oxidative stress to damage the mitochondrial structure.The dynamic-related protein 1（Drp-1）regulates the process of mitochondrial division.When mitochondria divide,Drp-1 is recruited to the mitochondria division site to format ring-shaped helical structures around splitting points,which can hydrolyze the GTP to release energy.Then,the structures cut the mitochondria,resulting in a series of reaction processes of mitochondrial damage.The aim of this study is to explore the role of Drp-1 in the damage of primary chicken hepatocytes injury induced by high copper.We used Mdivi-1 as inhibitor to inhibit the expression of Drp-1.The primary chicken hepatocytes cultured in vitro from embryo were served as cell models.The cells were divided into five groups,which are control group,copper treatment group I(50μM Cu²⁺),Cu²⁺+Mdivi-1 I group(50μM Cu²⁺+100μM Mdivi-1),copper treatment group II(100μM Cu²⁺),Cu²⁺+Mdivi-1II group(100μM Cu²⁺+100μM Mdivi-1),and control+Mdivi-1group.All groups were collected after incubating in carbon dioxide incubator for 24h.The changes of mitochondrial ATPase in different concentration groups were determined by ATPase detection technique.The mRNA expression of Drp-1,autophagy-related genes（P62,LC3-I,and LC3-II）,and apoptosis-related genes（CytC,Caspase3,and Bcl2）were detected by fluorescence quantitative PCR.Moreover,he activity of LDH,AST,ALT,TP,ALB,and GGT in cell culture media and the contents of SOD,CAT,MDA,and T-AOC in cells were measured.The results were as follows:（1）The ATP content in the copper-treated group was significantly lower than that in the control group,and the difference was extremely significant（P<0.05）.There was a significant difference between the copper treatment II group and the copper treatment I group（P<0.01）.After the inhibitor was added to inhibit the expression of Drp-1,the ATP content showed a certain upward trend.（2）Mdivi-1 effectively inhibited the expression of Drp-1,and,compared with the control,the copper-treated group produced significant difference in autophagy-related genes（P62,LC3-I,LC3-II）（P<0.01）.In terms of apoptotic genes,the copper-treated group succeeded in inducing the high expression of CytC and Caspase3,and the low expression of Bcl2（P<0.05）.（3）Compared with the control group,the activity of LDH,AST,ALT,and GGT in the copper-treated group was significantly higher（P<0.01）.All of them decreased after the inhibition of Drp-1 expression.The TP and ALB in the copper treatment group showed a significant decrease trend（P<0.01）.The inhibition of Drp-1 expression by the inhibitor Mdivi-1 could relieve the decrease of the TP and ALB.（4）The copper treatment group can significantly increase the content and activity of SOD,CAT,MDA,and T-AOC in the primary chicken hepatocytes（P<0.05）,but after inhibiting the expression of Drp-1,this increase trend was significantly alleviated（P<0.05）.Conclusion:Effectively inhibits the expression of Drp-1 can alleviate the oxidative damage induced by high copper.