Effects Of Allium Mongolicum Regel Flavonoids On Growth,antioxidant And Anti-inflammatory Activities Of Channa Argus

In recent years,aquatic animals have produced immune stress response due to environmental impact.In order to improve the immunity of aquatic animals and reduce the economic losses caused by immune stress,the development of green,safe and effective immune enhancers has become a hot issue.Allium mongolicum Regel flavonoids are organic compounds produced in Allium mongolicum,which have many physiological functions,such as improving the growth performance of mammals,regulating the body’s immune ability and alleviating cell damage.However,there is no report about AMRF in aquatic animals.Therefore,this experiment took Channa argus as the research object to study the effects of AMRF on the growth performance and immune function of Channa argus,and used lipopolysaccharide（LPS）as an immunostimulant to study the pre-protection effect of AMRF on LPS-induced inflammation.It provides a theoretical basis for making AMRF a new type of immune enhancer applied in aquaculture.The experiment is divided into four parts.（1）Effects of Allium mongolicum Regel flavonoids on growth,feed utilization and intestinal morphology of Channa argusIn this experiment,Channa argus with initial body weight of（3.44±0.31）g was fed with basic diet supplemented with 0（control group）,10,20,40 and 60 mg·kg^-1AMRF respectively for 8 weeks.The growth performance and intestinal morphology of Channa argus were measured.The results showed that,compared with the control group,20-60 mg·kg^-1AMRF could significantly improve the final body weight and feed efficiency of Channa argus（P<0.05）,40-60 mg·kg^-1AMRF could significantly improve the weight gain rate and specific growth rate of Channa argus（P<0.05）,10-60 mg·kg^-1AMRF had no significant effect on the liver to body ratio,viscera to body ratio and fatness of Channa argus（P>0.05）.In addition,when the AMRF dosage was 20-40 mg·kg^-1,the height of hindgut villi of Channa argus was significantly higher than that of the control group（P<0.05）,when the AMRF dosage was 20-60 mg·kg^-1,the thickness of hindgut muscle layer of Channa argus was significantly higher than that of the control group（P<0.05）.（2）Effects of Allium mongolicum Regel flavonoids on immunity and antioxidant capacity of Channa argusOn the basis of experiment 1,the serum and liver of Channa argus in each group were taken to determine the immune and antioxidant indexes.The results showed that,compared with the control group,the activities of superoxide dismutase（SOD）,glutathione peroxidase（GSH-PX）in serum and catalase（CAT）in liver of Channa argus were significantly increased by adding 10-60 mg·kg^-1AMRF（P<0.05）,20-60 mg·kg^-1AMRF significantly increased the activities of CAT in serum,SOD and GSH-PX in liver,and decreased the content of MDA in serum and liver（P<0.05）.Compared with the control group,10-60 mg·kg^-1AMRF significantly decreased the activities of AST and ALT,and increased the content of C3（P<0.05）,40-60mg·kg^-1AMRF significantly increased the levels of AKP,LZM and total protein（P<0.05）,20-60mg·kg^-1AMRF could significantly decrease the serum albumin content（P<0.05）,60 mg·kg^-1AMRF could significantly increase the serum immunoglobulin M（Ig M）content（P<0.05）,and there was no significant difference among the other groups,In addition,AMRF had no significant effect on the serum complement 4（C4）content（P>0.05）.（3）Allium mongolicum Regel flavonoids alleviates LPS induced inflammatory response in Channa argusOn the basis of experiment 2,the pre protective effect of AMRF on LPS induced inflammation in Channa argus was studied by intraperitoneal injection of LPS.0 mg·kg^-1PBS group was used as blank control group,and the other LPS injection groups were LPS+0 group,LPS+10 group,LPS+20 group,LPS+40 group and LPS+60 group.The results showed that compared with the blank control group,the activities of SOD,CAT and GSH-PX in serum and liver of Channa argus were significantly decreased,and the content of MDA was significantly increased（P<0.05）.Compared with LPS+0 group,10-60 mg·kg^-1AMRF significantly increased the activities of SOD and GSH-PX in serum and decreased the content of MDA in liver（P<0.05）,The addition of 20-60 mg·kg^-1AMRF significantly increased CAT activity in serum and liver,and decreased MDA content in serum（P<0.05）,Adding 40-60 mg·kg^-1AMRF could significantly increase the activities of SOD and GSH-PX（P<0.05）.Compared with the blank control group,LPS injection could significantly down regulate the levels of AKP,total protein,albumin,LZM,Ig M,C3 and C4,and up regulate the levels of AST and ALT（P<0.05）.Compared with LPS+0 group,adding 20-40 mg·kg^-1AMRF could significantly improve the AKP activity of Channa argus（P<0.05）,Adding 10-60 mg·kg^-1AMRF could significantly reduce the activity of AST and increase the content of C3（P<0.05）,The addition of 20-60mg·kg^-1AMRF significantly decreased the activity of serum ALT and increased the levels of serum albumin,LZM and C4（P<0.05）,Adding 40-60 mg·kg^-1AMRF could significantly increase the content of serum total protein（P<0.05）,Adding 10-40 mg·kg^-1AMRF could significantly increase the serum Ig M content（P<0.05）.Compared with the blank control group,the relative gene expressions of IL-1,IL-8,TNF-αand NF-κB p65 in liver,spleen,kidney and intestine of Channa argus after LPS injection were significantly increased（P<0.05）.Compared with LPS+0 group,10-60 mg·kg^-1AMRF could significantly reduce the gene expression levels of IL-1,IL-8,TNF-αand NF-κB p65 in liver,spleen,kidney and intestine（P<0.05）.（4）Effects of Allium mongolicum Regel flavonoids on blood lymphocyte activity,antioxidant capacity and relative gene expression of Channa argusIn this experiment,the effect of AMRF on blood lymphocytes of Channa argus was studied at the cellular level.The cell suspension was added with 0.00（control group）,25.00,50.00,100.00 and 200.00μg·m L^-1AMRF respectively.After 24 hours of culture,the cell activity,antioxidant index and gene expression level were determined.The results showed that compared with the control group,25-200μg·m L^-1AMRF could significantly improve the activity of blood lymphocytes（P<0.05）.50-200μg·m L^-1AMRF significantly increased the levels of T-AOC and nitric oxide（NO）in blood lymphocytes of Channa argus（P<0.05）,100-200μg·m L^-1AMRF significantly increased CAT activity（P<0.05）,250-200μg·m L^-1AMRF significantly increased the activities of T-SOD and GSH-PX（P<0.05）,100μg·m L^-1AMRF could significantly reduce the content of MDA（P>0.05）.The MDA levels of the other groups were lower than those of the control group,but the difference was not significant（P>0.05）.Compared with the control group,100μg·m L^-1AMRF could significantly reduce the expression of IL-1 and TNF-α,and increase the gene expression of HSP70,Hsp90 and IκBα（P<0.05）,50-200μg·m L^-1AMRF significantly decreased the expression of NF-κB p65 and increased the gene expression of glucocorticoid receptor（GR）（P<0.05）,25-200μg·m L^-1AMRF could significantly reduce IL-8 gene expression（P<0.05）.Under the experimental,AMRF can improve the growth performance,immune and antioxidant functions and stress resistance of Channa argus.According to the above indexes,In the diet of Channa argus,the optimal amount of added AMRF is 40-60 mg·kg^-1.In the blood lymphocytes of Channa argus,the optimal amount of added AMRF is 100-200μg·m L^-1.