Objective To analyze the relationship between serum levels of adrenocorticotropic hormone(ACTH)and thyroid hormone(TH)and intestinal flora in irritable bowel syndrome(IBS).Methods A retrospective study was conducted to select 178 patients diagnosed with IBS in our hospital from May 2021 to May 2023 as the observation group,and 216 healthy subjects as the control group.The number of typical bacterial flora in the intestine and its Alpha diversity(Chao1 index and Shannon index),the changes of ser-um levels of ACTH and TH,and the correlation between the two indicators and typical intestinal flora were respectively detected in the two groups.Results The bifidobacteria(Bifido)and lactobacillus(LaG-GG)in the observation group were lower than those in the control group,while the Escherichia coli(E.coli)and Salmonella(SE)in the observation group were higher than those in the control group(P<0.05).The Chaol index and Shannon index in the observation group were lower than those in the control group(P<0.05).The serum level of ACTH in the observation group was lower than that in the control group at 8:00(P<0.05),and higher than that in the control group at 24:00(P<0.05).There was no significant difference in the serum level of ACTH between the two groups at 16:00(P>0.05).The levels of T4 and FT3 in the observation group were higher than those in the control group(P<0.05),and the level of FT4 in the observation group was lower than that in the control group(P<0.05).There was no significant difference in the levels of T3 and TSH between the two groups(P>0.05).The levels of ACTH and FT4 in the observation group were positively correlated with Bifido and LaG-GG,and negative-ly correlated with E.coli and SE at 8:00(P<0.05).The levels of ACTH,T4 and FT3 were negatively correlated with Bifido and LaG-GG,and positively correlated with E.coli and SE at 24:00(P<0.05).Conclusion The diversity of intestinal flora in patients with IBS is low.The levels of ACTH and FT4 are positively correlated with Bifido and LaGGG,and negatively correlated with E.coli and SE at 8:00.The levels of ACTH,T4 and FT3 are negatively correlated with Bifido and LaG-GG,and positively correlated with E.coli and SE at 24:00.