The Effect Of Jujuboside On The Expression Of Feed Intake Related Genes In Hypothalamus Of Heat-stress Broiler

ObjectiveBy studying the effects of Jujuboside on AA broiler chickens hypothalamic NPY,GABAA-α1,Ag RP,MC4 R and POMC feed regulation factor expression,we proved the pathway of Jujuboside regulating feed intake in heat stressed Broilers.Furthermore,we disclosed the mechanism of Jujuboside improving heat stress broiler performance.MethodsThe samples of this experiment were 4-week-old AA+ broiler,which were feed up dietary Jujuboside(diets were added 0,1.0 mg/ kg Jujuboside),and6-week-old AA+broiler,which were suffered from heat stress in the project team(at normal temperature and 34 ℃)and were feed up dietary Jujuboside(diets were added 0,1.0 mg/kg Jujuboside).To detect mRNA and protein’s expression,which were from hypothalamic NPY,Ag RP,GABAA-α1,MC4 R and POMC genes,we used real-time PCR and Western Blotting.Then this study revealed the relationship between dietary jujuboside regulation and the broiler hypothalamus regulation gene’s expression with the broiler performance by using SPSS software for statistical analysis.Results1.Adding jujubosides could affect the hypothalamic feed intake regulationgene mRNA and proteins’ expression in 4 week old broiler.At the age of 4weeks,the NPY and GABAA-α1 mRNA level of male was extremely lower than female(P<0.01).While,there was no significant difference in Ag RP、MC4R and POMC mRNA between males and females(P > 0.05).When had added jujubosides diet,the males hypothalamic NPY、MC4R and POMC mRNA level were increased(P < 0.01)significantly.However,there was no significant change in the females(P > 0.05).At the age of 4 weeks,the females hypothalamic GABAA-α1 and POMC protein was significantly higher than males(P<0.01),but the females Ag RP protein was extremely lower than the males(P < 0.01).There were no significant differences in NPY and MC4 R protein.By adding jujubosides diet,Rooster hypothalamic NPY protein(P<0.01)and GABAA-α1 protein(P < 0.05)increased significantly,Hen hypothalamus Ag RP protein improved significantly(P<0.01).2.The effects of heat stress and add jujubosides hypothalamic feed intake regulation gene mRNA and proteins expression in 6 week old broiler.34 ℃heat stress significantly increased the chickens of hypothalamic Ag RP,MC4 R and NPY mRNA(P<0.01),POMC mRNA significantly lower(P<0.01).While,the males GABAA-α1 were extremely difference.Adding jujuboside to the fodder on heat stress of chicken made the hypothalamic Ag RP 、 MC4 R and NPY mRNA extremely decreased(P < 0.01 or P < 0.05).Heat stress causes the males hypothalamic GABAA-α1 、 MC4 R and POMC protein significantly decreased(P < 0.01),females hypothalamic NPY and POMC protein significantly decreased(P<0.01),However,the MC4 R protein increased(P<0.01).When add jujuboside in the fodder,we found that,the Ag RP 、GABAA-α1(males)、POMC(P<0.01)and NPY(P<0.05)protein in jujuboside group was extremely significant higher than heat stress group.While,the males hypothalamic MC4 R protein level was extremely significant increased(P <0.01).3.Heat stress and add the jujuboside to the fodder had influenced interacting relationship of ingestion related genes in the broiler chicken hypothalamus.6 weeks old rooster hypothalamic POMC protein and MC4 R protein(r=0.956,P=0.011),Ag RP protein(r=0.929,P=0.023),GABAA-α1protein(r=0.975,P=0.005)were positively correlated.The MC4 R protein and GABAA-α1 protein(r=0.881,P=0.048),Ag RP protein(r=0.968,P=0.007)were positively correlated.While,6 weeks old chicken hypothalamic MC4 R protein and Ag RP protein(r=0.710,P=0.021),GABAA-α1 protein(r=0.726,P=0.018)showed significantly positive correlation.The POMC protein and MC4 R protein(r=0.780,P=0.008),Ag RP protein(r=0.878,P=0.001),GABAA-α1protein(r=0.940,P=0.000)were positively correlated.The Ag RP protein and GABAA-α1 protein were positively correlated(r=0.686,P=0.029).4-6 weeks old chicken POMC protein respectively with GABAA-α1 protein(r=0.875,P=0.000)and MC4 R protein(r=0.647,P=0.012)showed significantly positive correlation.The MC4 R protein respectively with Ag RP protein(r=0.688,P=0.007)showed significantly positive correlation.Conclusions1.Jujuboside regulated NPY、Ag RP、MC4R、POMC、GABAA-α1 genes in4 weeks old and 6 weeks old heat stress chickens hypothalamus,and alleviated the negative effects that heat stress causing.2.At 6 weeks of age,Hen hypothalamus ADG and MC4 R protein showed significant negative correlation,while there was a positive correlation between NPY protein and ADG.At the age of 4-6 weeks,there was a positive correlation between ADFI and hypothalamus Ag RP protein,when ADG was significantly correlated with NPY protein.3.The best optical time that jujuboside was added to food was in heat stressing.