| The purpose of this study was to investigate the effects of acute dichlorvos exposure on cerebellar autophagy and cecal microflora in broilers,and to analyze the relationship between cerebellar autophagy related genes and cecal microflora.Thirty 40-day-old 817 broilers were randomly divided into three groups: control group(C Group),low dose group(L group)and high dose group(H Group).On the day of experiment,broilers in C group were given distilled water;broilers in L group were given 2.48 mg/kg of dichlorvos;broilers in H group were given 11.3 mg/kg of dichlorvos.After challenge,the samples of cecum and cerebellum were collected to detect the content or activity of SOD,cat,MDA and T-AOC in cerebellum.HE staining sections of cerebellum and cecum and LC3 immunofluorescence sections of cerebellum were made.RT-PCR and Western blot were used The expression of genes related to oxidative stress and autophagy in cerebellum was detected by blot.The contents of cecum were analyzed by 16 S r RNA high-throughput sequencing.Finally,the correlation between the expression of autophagy-related genes and intestinal microorganism was analyze.1.Compared with the C group,the cerebellar sulcus space became lager,glial cell deformation,Purkinje cell vacuole deformation,granular cell shrinkage and deformation appeared in the two groups;the cecum of the two groups showed obvious thickening of muscle layer,shortening or breaking of intestinal villi,atrophy of intestinal gland and reduction of goblet cell number.2.Compared with the C group,the activities of SOD and T-AOC were significantly increased(P<0.05 or P<0.01),and the content of MDA was significantly increased(P<0.01);in addition,the mRNA expression levels of Nrf2,HO-1,SOD-1,cat,GSH,GCLC,GCLM and NQO1 were significantly increased in the L group(P<0.05 or P<0.01);the mRNA expression levels of SOD-1,GSH,GCLC,GCLM and NQO1 in the H group were significantly increased(P<0.05 or P<0.01).3.Compared with the C group,the mRNA expression levels of LC3-A,LC3-B and Atg5 in L group were significantly up-regulated(P<0.001 or P<0.01),and the mRNA expression levels of m TOR were significantly down regulated(P<0.01);the mRNA expression levels of LC3-A,LC3-B and Beclin1 in H group were significantly up-regulated(P<0.05 or P<0.01),and the mRNA expression levels of P62 and m TOR were significantly down regulated(P<0.05 or P<0.01).Protein level results showed that LC3-II/LC3-I protein expression was significantly up-regulated in low-dose group(P<0.01),the expression of Beclin1 protein in high-dose group was significantly increased(P<0.05).4.Compared with the C group,at the phylum level,the relative abundance of Cyanobacteria in the L group increased significantly(P<0.01),while the relative abundance of 9 phylum,such as Methylomirabilota,Nitrospirota and Armimonadota,decreased significantly(P<0.05 or P<0.01),and the relative abundance of Fusobacteria in the H group decreased significantly(P<0.05).At the genus level,the relative abundances of 4 genera,such as unidentified chloroplast,unidentified mitochondria and Entercoccus in L group increased significantly(P<0.05 or P<0.01),while the relative abundances of 20 genera,such as Porphyromonas,Prevotella and Neisseria in L group decreased significantly(P<0.05 or P<0.01).The relative abundances of 9 genera,such as Barnesiella,Acinetobacter and Flavonifractor,were significantly increased(P<0.05 or P<0.01),while those of 7 genera,such as Porphyromonas,Prevotella and Neisseria,were significantly decreased(P<0.05 or P<0.01)in the H group.5.The up regulation or down regulation of autophagy related gene mRNA expression is related to specific bacteria in cecum of broilers.In conclusion,acute dichlorvos exposure damaged the normal morphological structure of the cerebellum and cecum of broilers,caused pathological injury of the cerebellum and cecum,induced oxidative stress and autophagy in the cerebellum of broilers,caused cerebellum injury,and reduced the diversity of cecal flora and destroyed the stability of cecal microflora. |
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