The Role Of Vitamin A In Gastrointestinal Symptoms,gut Microbiotaand Behaviors In Children With Autism Spectrum Disorder

PART I THE RELATIONSHIP BETWEEN GASTROINTESTINAL SYMPTOMS,GUT MICROBIOTA AND BEHAVIORS IN CHILDREN WITH AUTISM SPECTRUM DISORDERBackground: Gastrointestinal(GI)symptoms and dysbiosis of gut microbiota are commonly reported in autism spectrum disorder(ASD),which may contribute to behavioral impairment via gut-brain axis.Objective: To find the GI symptoms in ASD and analyze the relationship between GI symptoms and behaviors.To find the related gut microbiota in ASD without GI symptoms.To find the most common GI symptom in ASD,analyze the damage of behaviors in this subtype and related gut microbiota.Methods: The autism behaviour checklist(ABC),childhood autism rating scale(CARS)and social responsiveness scale(SRS)were used to assess autism symptoms.CD38 and acid-related orphan receptor alpha(RORA)mRNA levels were used to assess autism behavior related biochemical indicators' changes,6-GI severity index(6-GSI)was used to assess GI symptoms.183 children with ASD and 66 typical developmental children participated in this study,completed all above.The difference of GI symptoms between ASD and CON,the relationship between GI symptoms and ABC,CARS,SRS,CD38,RORA mRNA levels in ASD were analyzed.The difference of ABC,CARS,SRS,CD38,RORA mRNA levels were evaluated between the subtype of ASD with most common GI symptom and ASD without GI symptom.Illumina MiSeq for 16 S rRNA genes was used to compare the differences in gut microbiota between ASD-GI and ASD+most common GI symptom,ASD-GI and CON-GI in the subsets,with each group of 24,randomly selected from the ASD children who finished the stool collection.Results: The prevalence of total GI symptoms(52.46% VS.36.36%)and constipation(33.33% VS.15.15%)were significantly more common in ASD than CON(all P < 0.05).After adjusting the confounding factors,the GI socres(1.08 ± 1.44 VS.0.68 ± 0.98)and constipation scores(0.40 ±0.62 VS.0.17 ± 0.41)were significantly higher in ASD than CON(all adjust P < 0.05).In children with ASD,GI scores were not significantly correlated with ABC,CARS,SRS scores(all P > 0.05);RORA mRNA level were significantly negatively correlated with total GI score(?s=-0.21,P=0.0053)and constipation(?s=-0.18,P=0.0184);CD38 mRNA level werenot significantly correlated with GI scores(all P >0.05).There were no significant differences in ABC,CARS,SRS scores between ASD with constipation and ASD without GI symptoms(all P >0.05),neither in CD38 mRNA level(P >0.05).RORA mRNA level was significantly lower in ASD with constipation than ASD without GI symptoms(P=0.0075).In the gut microbiota analysis subgroups,ASD-GI were enriched in Prevotella,Bifidobacterium,Lactobacillales,Streptococcus,Erysipelotrichaceae,etc than CON-GI in LEfSE analysis,while were enriched in Coriobacteriia,Coriobacteriales,Erysipelotrichales,Pseudomonadales than CON-GI in Wilcoxon analysis.ASD+constipation were enriched in Prevotellaceae,Prevotella,Paraprevotella,Megasphaera,Christensenellaceae and lowered in Eggerthella than ASD-GI in LEfSE analysis.But the significant differences between ASD+constipation and ASD-GI can not be repeated in Wilcoxon analysis.Conclusions: GI symptoms were more common in ASD than CON;GI symptoms positively correlated with social behavior related marker damage and Lactobacillales,Streptococcus,Bifidobacterium significantly increased in ASD-GI.Constipation was the most common GI symptom in ASD and ASD with constipation may have more damage in social behavior related marker.Constipation related gut micriobiota were not foud,it's open to futher study.PART ? RELATIONSHIP BETWEEN VITAMIN AAND GASTROINTESTINAL SYMPTOMS,GUT MICROBIOTA,BEHAVIORS IN CHILDREN WITH AUTISM SPECTRUM DISORDERBackground: Our preliminary work showed that GI symptoms were common in ASD,along with gut microbiota changes,which may contribute to autism behavior related biochemical marker changes.Vitamin A(VA)plays a role in regulation of both central nervous system(CNS)development and intestinal mechanical barrier,immune barrier,biological barrier.Objective: This study was performed to investigate the role of VA in the changes of GI symptoms,gut microbiota and behaviors in children with ASD.Methods: High performance liquid chromatography(HPLC)was used to assess plasma retinol levels.ABC,CARS and SRS were used to assess autism symptoms.CD38 and RORA m RNA levels were used to assess autism behavior related biochemical indicators' changes.6-GSI was used to assess GI symptoms.Fisrt,all ASD participants from part I accepted tests about plasma retinol,GI symptoms,ABC,CARS,SRS,CD38 and RORA m RNA levels.Relationship between VA levels and GI scores,VA levels and autism symptoms,VA levels and biochemical indicators were analyzed.Second,sixty four children with ASD completed a 6-month follow-up study with VA intervention.Evaluations of plasma retinol,GI symptoms,ABC,CARS,SRS,CD38 and RORA m RNA levels were performed before and after 6 months of intervention in the 64 children.Illumina Mi Seq for16 S r RNA genes was used to compare the differences in gut microbiota before and after 6 months of treatment in the subset 20 of the 64 children.Results: In the 64 autistic children,VA were significantly negatively correlated with total GI scores,stool consistency scores(?s=-0.15,?s=-0.18,P=0.0449,P=0.0268);VA were not significantly correlated with ABC,CARS,SRS scores(all P > 0.05);VA were significantly correlated with CD38 and RORA m RNA levels(?s=0.30,?s=0.21,P=0.0001,P=0.0097).After 6 months of intervention,the total GI problems,constipation,diarrhea,stool smell scores significantly decreased(all P < 0.05);plasma retinol,CD38 and RORA m RNA levels significantly increased(all P <0.05);the scores of ABC,CARS and SRS scales showed no significant differences(all P > 0.05).Meanwhile,in the subgroup of 20,after 6 months of intervention,the proportion of Bacteroidetes/Bacteroidia/Bacteroidales(phylum to order level)significantly increased and the proportion of Actinobacteria/Bifidobacteriales/Bifidobacteriaceae/Bifidobacterium(class to genus level)significantly decreased no matter using LEf SE or Wilcoxon tests(all P< 0.05).Conclusions: Bacteroidetes/Bacteroidales maybe the key taxa related to VA.Moreover,VA played a role in the changes in GI symptoms and autism social behavior related biomarkers.It remains unclear whether the VA concentration is associated with autism symptoms.