基于16S rDNA探讨芎麻汤有效成分对偏头痛大鼠肠道微生物的影响OA

Objective To study the effect of Xiongmatang active ingredients on intestinal microbes in a rat migraine model.Methods One hundred and twenty rats underwent dural cannulation surgery,and general physical signs were observed.Ninety healthy rats were selected and injected with inflammation soup six times within 12 days to stimulate the dura mater to create a migraine model.These rats were randomly divided into nine groups of 10 rats:model,calcitonin gene-related peptide(CGRP)inhibitor,flunarizine-positive(Flunarizine),and Xiongmatang active ingredients(low-and high-dose groups for n-butanol,ethyl acetate,and n-butanol+ethyl acetate extracts).An additional control group of 10 rats was established,injected with 0.9%sodium chloride solution using the same modeling procedure.After successful modeling,the control and model groups were given pure water by gavage,and the CGRP inhibitor group were given the drug by tail vein five times,once a week;the other groups were given corresponding doses,once a day,for a total of 4 weeks(28 d).Behavioral assessments were then conducted using sucrose preference,tail suspension,novel object recognition,and Morris water-maze tests.Abdominal aortic blood was collected for enzyme-linked immunosorbent assay quantification of circulating CGRP and NO levels.Intestinal contents were taken,and the changes in the gut microbiota were analyzed by 16S rDNA high-throughput sequencing.Results Compared with the control group,model group rats exhibited significant depressive-like behavioral changes and cognitive impairments,meanwhile,the contents of CGRP and NO in serum were significantly increased(P<0.05).Gut microbiota analysis revealed alterations in microbial community structure.At the phylum level,the abundance of Firmicutes,Actinobacteriota,Unclassified,and Campylobacterota decreased significantly(P<0.01,P<0.05),while that of Bacteroidota,Verrucomicrobiota,Patescibacteria,and Cyanobacteria increased significantly(P<0.01,P<0.05).At the genus level,abundances of certain genera,such as Muribaculaceae_unclassified and Lactobacillus,were significantly decreased(P<0.05),while Ruminococcus,Clostridia_UCG-014_unclassified,Akkermansia,Firmicutes_unclassified,and Monoglobus significantly increased(P<0.01,P<0.05).Compared with the model group,the CGRP inhibitor,Flunarizine group,and Xiongmatang active ingredients dose-dependently reversed these depressive-like behavioral changes and cognitive impairments,and significantly lowered serum levels of CGRP and NO.The active ingredients of Xiongmatang treatment also modulated intestinal microbial diversity;α-diversity assays showed increased richness and evenness,while β-diversity revealed that the microbial community structure of Xiongmatang-treated rats closely resembled that of the control group.At both phylum and genus levels,the alterations in microbiota composition exhibited trends opposite to those seen in the model group.Conclusions The active ingredients of Xiongmatang may prevent and treat migraines in rats through gut microbiota regulation.