Objective:

2

To explore the possible effects and mechanism of Zhizhu Decoction (ZZD) on the pathophysiology of slow transit constipation (STC).

Methods:

2

A total of 54 C57BL/6 mice was randomly divided into the following 6 groups by a random number table

including control

STC model (model)

positive control

and low-

medium- and high-doses ZZD treatment groups (5

10

20 g/kg

namely L

M-

and H-ZZD

respectively)

9 mice in each group. Following 2-week treatment

intestinal transport rate (ITR) and fecal water content were determined

and blood and colon tissue samples were collected. Hematoxylin-eosin and periodic acid-Schiff staining were performed to evaluate the morphology of colon tissues and calculate the number of goblet cells. To determine intestinal permeability

serum levels of lipopolysaccharide (LPS)

low-density lipoprotein (LDL) and mannose were measured using enzyme-linked immunosorbent assay (ELISA). Western blot analysis was carried out to detect the expression levels of intestinal tight junction proteins zona-occludens-1 (ZO-1)

claudin-1

occludin and recombinant mucin 2 (MUC2). The mRNA expression levels of inflammatory cytokines including tumor necrosis factor (TNF)-α

interleukin (IL)-1β

IL-6

IL-4

IL-10 and IL-22 were determined using reverse transcription-quantitative reverse transcription reaction. Colon indexes of oxidative stress were measured by ELISA

and protein expression levels of colon silent information regulator 1/forkhead box O transcription factor 1 (SIRT1/FoxO1) antioxidant signaling pathway were detected by Western blot.

Results:

2

Compared with the model group

ITR and fecal moisture were significantly enhanced in STC mice in the M-ZZD and H-ZZD groups (

P

<

0.01). Additionally

ZZD treatment notably increased the thickness of mucosal and muscular tissue

elevated the number of goblet cells in the colon of STC mice

reduced the secretion levels of LPS

LDL and mannose

and upregulated ZO-1

claudin-1

occludin and MUC2 expressions in the colon in a dose-dependent manner

compared with the model group (

P

<

0.05 or

P

<

0.01). In addition

ZZD significantly attenuated intestinal inflammation and oxidative stress and activated the SIRT1/FoxO1 signaling pathway (

P

<

0.05 or

P

<

0.01).

Conclusion:

2

ZZD exhibited beneficial effects on the intestinal system of STC mice and alleviated intestinal inflammation and oxidative stress via activating SIRT1/FoxO1 antioxidant signaling pathway in the colon.