Objective:

2

To investigate whether

Radix Sanguisorbae

(RS

Diyu) could restore intestinal barrier function following sepsis using a cecal ligation and puncture (CLP)-induced septic rat model and lipopolysaccharide (LPS)-challenged IEC-6 cell model

respectively.

Methods:

2

Totally 224 rats were divided into 4 groups including a control

sham

CLP and RS group according to a random number table. The rats in the control group were administrated with Ringer's lactate solution (30 mL/kg) with additional dopamine [10 μg/(kg • min)

]

and given intramuscular injections of cefuroxime sodium (10 mg/kg) 12 h following CLP. The rats in the RS group were administrated with RS (10 mg/kg) through tail vein 1 h before CLP and treated with RS (10 mg/kg) 12 h following CLP. The rats in the sham group were only performed abdominal surgery without CLP. The rats in the CLP group were performed with CLP without any treatment. The other steps were same as control group. The effects of RS on intestinal barrier function

mesenteric microvessels barrier function

multi-organ function indicators

inflammatory response and 72 h survival window following sepsis were observed.

In vitro

the effects of RS on LPS-challenged IEC-6 cell viability

the expressions of zona occludens-1 (ZO-1) and ferroptosis index were evaluated by cell counting kit-8

immunofluorescence and Western blot analysis. Bioinformatic tools were applied to investigate the pharmacological network of RS in

sepsis to predict the active compounds and potential protein targets and pathways.

Results:

2

The sepsis caused severe intestinal barrier dysfunction

multi-organ injury

lipid peroxidation accumulation

and ferroptosis

in vivo

. RS treatment significantly prolonged the survival time to 56 h and increased 72-h survival rate to 7/16 (43.75%). RS also improved intestinal barrier function and relieved intestinal inflammation. Moreover

RS significantly decreased lipid peroxidation and inhibited ferroptosis (

P

<

0.05 or

P

<

0.01). Administration of RS significantly worked better than Ringer's solution used alone. Using network pharmacology prediction

we found that ferroptosis and hypoxia inducible factor-1α (HIF-1α) signaling pathways might be involved in RS effects on sepsis. Subsequent Western blot

ferrous iron measurements

and FerroOrange fluorescence of ferrous iron verified the network pharmacology predictions.

Conclusion:

2

RS improved the intestinal barrier function and alleviated intestinal injury by inhibiting ferroptosis

which was related in part to HIF-1α/heme oxygenase-1/Fe

2+

axis.