Objective:

2

To explore the mechanism of colon dialysis with Yishen Decoction (YS) in improving the autophagy d

isorder of intestinal epithelial cells in chronic renal failure (CRF)

in vivo

and

in vitro

.

Methods:

2

Thirty male SD rats were randomly divided into normal

CRF

and colonic dialysis with YS groups by a random number table method (

n

=10). The CRF model was established by orally gavage of adenine 200 mg/(kg•d) for 4 weeks. CRF rats in the YS group were treated with colonic dialysis using YS 20 g/(kg•d) for 14 consecutive days. The serum creatinine (SCr) and urea nitrogen (BUN) levels were detected by enzyme-linked immunosorbent assay. Pathological changes of kidney and colon tissues were observed by hematoxylin and eosin staining. Autophagosome changes in colonic epithelial cells was observed with electron microscopy.

In vitro

experiments

human colon cancer epithelial cells (T84) were cultured and divided into normal

urea model (74U)

YS colon dialysis

autophagy activator rapamycin (Ra)

autophagy inhibitor 3-methyladenine (3-MA)

and SIRT1 activator resveratrol (Re) groups. RT-PCR and Western blot were used to detect the mRNA and protein expressions of zonula occludens-1 (ZO-1)

Claudin-1

silent information regulator sirtuin 1 (SIRT1)

LC3

and Beclin-1 both

in vitro

and

in vivo

.

Results:

2

Colonic dialysis with YS decreased SCr and BUN levels in CRF rats (

P

<

0.05)

and alleviated the pathological changes of renal and colon tissues. Expressions of SIRT1

ZO-1

Claudin-1

Beclin-1

and LC3Ⅱ/Ⅰ were increased in the YS group compared with the CRF group

in vivo

(

P

<

0.05). In

in vitro

study

compared with normal group

the expressions of SIRT1

ZO-1

and Claudin-1 were decreased

and expressions of Beclin-1

and LC3Ⅱ/Ⅰwere increased in the 74U group (

P

<

0.05). Compared with the 74U group

expressions of SIRT1

ZO-1

and Claudin-1 were increased

whereas Beclin-1

and

LC3Ⅱ/Ⅰwere decreased in the YS group (

P

<

0.05). The treatment of 3-MA and rapamycin regulated autophagy and the expression of SIRT1. SIRT1 activator intervention up-regulated autophagy as well as the expressions of ZO-1 and Claudin-1 compared with the 74U group (

P

<

0.05).

Conclusion:

2

Colonic dialysis with YS could improve autophagy disorder and repair CRF intestinal mucosal barrier injury by regulating SIRT1 expression in intestinal epithelial cells.