Objective:

2

To explore whether polysaccharide of Danggui Buxue Decoction (formula polysaccharide

FP) could attenuate colorectal cancer (CRC) via modulating intestinal microflora and metabolites.

Methods:

2

A total of 30 male C57BL/6 mice were randomly assigned to 3 groups according to body weight

including normal control

CRC model and FP groups (

n

=10). Dextran sulfate sodium and azoxymethane were used to induce CRC model. The mice in FP group were treated with FP [0.9 g/(kg•d)

]

for 8 weeks. Then the

coloretal length

weight and tumors number of colorectal tissues were observed. Colonic tissues were stained with hematoxylin and eosin to evaluate the histological changes. Tight junction proteins including claudin 1 and zonula occludens-1 (ZO-1) were detected using immunohistochemistry. Serum concentrations of endothelial cell specific molecule-1 and carcino embryonic antigen were determined using enzyme-linked immunosorbent assays. Toll-like receptor 4 (TLR4)/nuclear transcription factor-κB (NF-κB) p65 signal-related mRNA and proteins expressions were detected using real-time polymerase chain reaction and Western blot

respectively. Feces samples were detected with 16S rRNA and liquid chromatography-mass spectrometry and non-targeted metabolomics sequencing was used to analyze the composition of intestinal microbiota and metabolic changes.

Results:

2

FP significantly inhibited colorectal tumor growth

attenuated body weight loss

repaired colonic structure

improved intestinal barrier dysfunction

reduced colonic inflammatory cytokine levels and inhibited TLR4/NF-κB pathway related mRNA and protein expressions (all

P

<

0.01). Moreover

FP altered the gut microbiota composition of CRC mice dramatically

characterized by a reduction of

Firmicutes

-

Bacteroidetes

ratio at the

phylum

level. FP suppressed

Lachnospiraceae

_NK4A136_group

Odoribacter

and

Romboutsia

(

P

<

0.05 or

P

<

0.01)

and elevated the abundance of

Dubosiella

Candidatus

_

Saccharimonas

and

Alloprevotella

at the

genus

level (

P

<

0.01). Non-targeted metabolomics sequencing detected significant differences in metabolites

and the functions of differential metabolites were focused on regulating amino acid metabolism

lipid metabolism and metabolism of cofactors and vitami

ns.

Conclusions:

2

FP attenuates colonic injury and intestinal inflammatory response in CRC mice

repairs disrupted-intestinal microbiota

and improves metabolites. This research provides experimental basis for application of FP as a potential therapeutic agent for CRC.