The Protective Effects And Mechanism Of Flavonoids From Allium Mongolicum Regel On Liver In Oxidative Stressed Small Tail Han Sheep

The present study was to examine the protective effect and its mechanism of Allium Mongolicum Regel(AMR)and flavonoids from Allium Mongolicum Regel(FAMR)on oxidative stress in Diquat-induced Small Tail Han Sheep.The sheep were used as an experimental animal model to study the effects of AMR and FAMR on antioxidant enzymes,antioxidants,mitochondrial function,and mitochondrial respiratory chain complex related-gene expression in the liver.This study consists of 4 experiments:Experiment 1:Effects of AMR and FAMR on antioxidant index in liver of Diquat-induced Small Tail Han Sheep.Fifteen Small Tail Han Sheep(body weight 40±5 kg)were randomly divided into three groups:control group(n=5),AMR group(n=5)and FAMR group(n=5).At the first day of the experiment,the control group was fed with basal diet,and the AMR group and FAMR group were fed with 10 g/d/sheep AMR and 120 mg/d/sheep FAMR.Respectively,on the 30th day of the experiment,Diquat solution was injected intraperitoneally at 10 mg/kg BW for 12 days.On the 42nd days,three sheep in each group were randomly selected to take liver tissue after slaughtering.The activities of SOD,CAT,GSH-Px,XOD,the content of MDA,GSH,T-AOC in the liver were measured.The results showed that feeding AMR and FAMR may protect the oxidative damage caused by Diquat to Small Tail Han Sheep by increasing the activity of SOD,CAT,GSH-Px and the content of GSH,and by inhibiting the activity of XOD and by decreasing the production of MDA.The protecting effect of FAMR was significantly higher than that of AMR(P<0.05).Experiment 2:Effects of AMR and FAMR on mitochondrial function in liver of Diquat-induced Small Tail Han Sheep.Based on the result of experiment 1,this experiment used molecular biology methods to explore the mechanism of effect of AMR and FAMR on the liver anti-oxidative stress from the perspective of mitochondrial function.The activities of mitochondrial respiratory chain complex ?,?,and mitochondrial ATPase were quantitatively detected by colorimetric method;the content of ROS in mitochondria was determined by fluorescence spectrophotometry;and the mitochondrial membrane potential was detected by flow cytometry.The results showed that feeding AMR and FAMR by lowering Diquat-induced Small Tail Han Sheep mitochondrial ROS levels in the liver,may improve the liver mitochondrial ATPase,mitochondrial respiratory chain complexes ? activity,and the level of the mitochondrial membrane potential.The protective effect of FAMR was significantly greater than that of AMR(P<0.05).Experiment 3:Effects of AMR and FAMR on the expression of the genes related to mitochondrial respiratory chain complex in Diquat-induced Small Tail Han Sheep.Reverse Transcription Polymerase Chain Reaction(RT-PCR)was used to investigate the effect of AMR and FAMR on the expression of SDH-related genes(SDHA,SDHB,SDI IC and SDHD)in the liver.The results showed that expression of SDHA in AMR group and FAMR group was significantly greater than that in control group(P<0.05),and it's expression in FAMR group was significantly greater than that in AMR group(P<0.05).The expression of SDHB,SDHC,and SDHD genes in flavonoids group was significantly greater than that in control group and that in AMR group(P<0.05).Overall,the oxidative damage induced by Diquat in Small Tail Han Sheep might be protected by feeding AMR and FAMR.The underling mechanism might be associated with the elimination of free radicals in the body,maintaining the balance of antioxidant enzymes,and antioxidants in vivo and protecting the function of mitochondria.