Objective

2

To explore the method of establishing a disease-syndrome combined model of irritable bowel syndrome （IBS-D） with spleen and kidney

yang

deficiency in mice.

Methods

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Model establishment： Twenty mice were randomly divided into a normal group and a model group， with 10 mice in each group. The model group was given 50 mg/（kg·d） of 0℃ ice adenine suspension by gavage for 14 days， plus tail-clamping stimulation and restrained in centrifuge tubes for 7 days， and 10 g/（kg·d） of 0 ℃ ice senna decoction by gavage for 5 days to prepare IBS-D spleen and kidney

yang

deficiency syndrome model. The normal group was given 0.4 ml of sterile water by gavage once a day for 14 days. Behavioral characteristics， food intake， fecal water content， body weight， and rectal temperature were observed in both groups. Pain threshold， gastric residual rate， intestinal propulsion rate， and the content of short-chain fatty acids （SCFAs）， serotonin （5-HT）， adrenocorti

cotropic hormone （ACTH）， corticosterone （CORT）， and D-xylose in the serum were detected after modeling. Model validation： Forty mice were randomly divided into a blank group （n=10） and a modeling group （n=30）. After successful modeling using the above method， the modeling group was divided into a model control group，

Sishen Pill

（四神丸） group， and pinaverium bromide group， with 10 mice in each group. The

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group was given 5 g/（kg·d） of

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decoction by gavage， and the pinaverium bromide group was given 21.63 mg/（kg·d） of pinaverium bromide solution by gavage， while the blank group and the model control group were given 0.70 ml/d of sterile water by gavage， all for 7 days. The indicators were detected as the same with model establishment.

Results

2

Mice in the model group had poor mental status， lethargy， dull hair and loose feces. Compared with the normal group， the model group exhibited reduced food intake and increased fecal water content ； on the 14th day of modeling， the model group showed a slower body weight gain rate and decreased rectal tempe-rature； after 14 days， the model group had increased small intestinal propulsion rate， and serum SCFAs and 5-HT levels with reduced serum levels of ACTH， CORT， and D-xylose （

P

＜0.05 or

P

＜0.01）. Model validation indicated that the

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group showed improvements in mental state， activity levels， fur smoothness and curling and gathering symptoms， while these symptoms in the pinaverium bromide group were not significantly improved. Compared to the model control group， the

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group and pinaverium bromide group had reduced fecal water content and increased food intake， as well as increased body weight gain and elevated rectal temperature on day 4. On the 7th day of administration， the pinaverium bromide group showed lower rectal temperature than

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group， and

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group showed decreased serum SCFAs and 5-HT levels with increased ACTH， CORT， and D-xylose levels （

P

＜0.05 or

P

＜0.01）， while the pinaverium bromide group exhibited reduced 5-HT and elevated CORT level （

P

＜0.01）.

Conclusion

2

A combination of ice adenine plus ice senna leaf gavage， tail clamping， and centrifuge tube restraint can successfully establish a disease-syndrome combined mouse model of IBS-D with spleen-kidney Yang deficiency.