The Relieving Effects And Mechanism Of Allium Mongolicum Regel And Its Flavonoids On Small Tail Han Sheep In Stress Condition Induced By Diquat

In this paper,Small Tail Han Sheep fed with Allium Mongolicum Regel(AMR)or Flavonoids from Allium Mongolicum Regel(FAMR)was induced a chronic oxidative stress model by Diquat,to explore AMR and FAMR whether have mitigation in the disease or not.It can provide a theoretical basis for the development and utilization of AMR as an antioxidant.The main contents and results of this thesis are as follows:Experiment 1:Establishment of a chronic oxidative stress model of Small Tail Han Sheep.Twelve Small Tail Han Sheep(body weight 35 ±5 kg)were randomly divided into four groups:control group(equal amount of normal saline,n=3),the low concentration group(8 mg/kg BW Diquat,n=3),the medium concentration group(10 mg/kg BW Diquat,n=3)and the high concentration group(12 mg/kg BW Diquat,n=3),respectively.Recording feed intake of sheep every day in the trial period.Recording weight and collecting venous blood at 0,6,12,18,24 d were conducted before morning diet.The determination of serum Superoxide Dismutase(SOD),Catalase(CAT),Glutathione Peroxidase(GSH-Px),Total antioxidant capacity(T-AOC)and Malondialdehyde(MDA),and other antioxidant index were examined.The results showed that the average daily intake(ADFI),average daily gain(ADG),serum SOD,CAT,GSH-Px activity and T-AOC in every Diquat concentration groups were significantly decreased(P<0.05),and MDA content was significantly increased(P<0.05),compared with the control group.On the 12th day of the experiment,compared with the control group,the serum antioxidant indexes,ADFI and ADG of sheep in the Diquat low concentration group were not significantly difference(P>0.05).It demonstrate that the duration of oxidative stress caused by Diquat low concentration was short.Sheep in the Diquat high concentration group died on the 6th day after the injection of Diquat,demonstrating that the oxidative stress generated by this concentration was too strong to be used in the follow-up study.On the 6th day,In the Diquat medium concentration group,compared with the control group,the activity of SOD,CAT,GSH-Px and T-AOC were significantly decreased(P<0.05),and MDA content significantly increased(P<0.05).On the 12th day,all the indicators reached the peak,while on the 18th day,despite the indicators recovered,they were still significantly lower than the control group(P<0.05).It indicates that the peritoneal injection of 10 mg/kg BW Diquat solution can maintain the oxidative stress of Small Tail Han Sheep for at least 18 days.The medium concentration of Diquat achieve the purpose of generating chronic oxidative stress.Therefore,10 mg/kg BW Diquat was selected for the subsequent experimental study.Experiment 2:The alleviation effect of Mongolicum Regel and Flavonoids from Allium Mongolicum Regel on Diquat-induced oxidative stress of 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.During the whole experimental period,the samples were weighed and collected before morning feeding at 0,15,30,34,38 and 42 d.The serum was separated and the antioxidant indexes(SOD,CAT,GSH-Px,T-AOC and MDA)in the serum were determined.On the 30th day of the experiment(before Diquat injection),compared with the control group,the results showed that ADFI,ADG,SOD,GSH-Px,CAT and T-AOC were significantly increased(P<0.05),and MDA content was significantly decreased(P<0.05)in AMR group and FAMR group.After the injection of Diquat,the indicators of AMR group and FAMR group were better than the control group,and the FAMR group was better than AMR group.The results showed that AMR group and FAMR group could alleviate the oxidative stress induced by Diquat,and the relief effect of FAMR was better than AMR.Experiment 3:Effects of Allium Mongolicum Regel and Flavonoids from Allium Mongolicum Regel on diquat-induced oxidative stress antioxidant mechanism in Small Tail Han Sheep.The experiment design is the same as experiment 2.The expression levels of NF-?B and iNOS in the liver of Small Tail Han Sheep were determined by reverse transcription polymerase chain reaction(RT-PCR).Compared with the control group,the results showed that AMR and FAMR could significantly inhibit the expression of NF-?B and iNOS genes(P<0.05)and reduce the production of NO.The results showed that the inhibition of NF-?B and iNOS gene expression in the liver of Diquat-induced Small Tail Han Sheep could reduce the production of NO.In conclusion,10 mg/kg BW Diquat solution can establish a chronic model of oxidative stress.The oxidative damage induced by Diquat in Small Tail Han Sheep might be alleviated by supplement AMR and FAMR in diet.The effect of FAMR is significantly better than AMR.