| Objective: To observe the effect of Shu-gan-jian-pi Decoction on the body weight growth,visceral sensitivity change,brain gut peptide(5-HT,VIP)and inflammatory cytokines(IFN-?,IL-10)in serum of IBS-D rats with liver depression and spleen deficiency,so as to explore the mechanism of Shu-gan-jian-pi decoction in the treatment of IBS-D and provide a new theoretical basis for the treatment of diarrhea type irritable bowel syndrome with traditional Chinese medicine.Materials and methods:In this experiment,48 healthy SPF SD rats 180-220 g were selected,half male and half female.After one week of adaptive feeding,the rats were randomly divided into two groups:blank group(12 rats)and model group(36 rats).The model rats were made by chronic restraint stress,tail clamping stimulation and senna leaf perfusion.Modeling method: chronic restraint stress,3 hours/day;tail clamping stimulation,half an hour/day;senna leaf,0.45g/ml,10ml/kg,once a day,14 days.After modeling,the rats in the model group were randomly divided into model control group,Shu-gan-jian-pi decoction group,and Siliankang group,with 12 rats in each group.At the end of modeling: 1.Observe the general situation of rats;2.The visceral sensitivity and diarrhea index of rats were detected by abdominal wall reflex(AWR)to evaluate the disease characteristics;3.Open field test,sugar water preference and tail suspension test were selected to evaluate the syndrome characteristics of liver depression;4.The weight gain on the 7th and 14 th day and the food intake within one week were selected to evaluate the syndrome characteristics of spleen deficiency;5.HE staining sections were prepared to observe the pathological changes and evaluate the success of the model.Treatment period: Rats in the blank group were not treated specially,and model control group were fed with normal saline once a day for 14 consecutive days;The rats in the Shugan Jianpi recipe group were treated with traditional Chinese medicine decoction with a concentration of2.0475 g/ml and a dosage of 10 ml/kg once a day for 14 consecutive days;The rats in Siliankang group were treated with Siliankang tablet diluted suspension by intragastric administration,the concentration was 0.04725g/ml,and the dosage was 10ml/kg once a day for 14 consecutive days.After treatment: 6.Observe the general situation of rats;7.Observethe weight change of rats;8.Observe the abdominal wall reflex(AWR)to detect the visceral sensitivity of rats;9.The content of brain-gut peptide(5-HT,VIP)in serum of rats in each group was detected by enzyme-linked immunosorbent assay;10.The changes of inflammatory cytokines(IFN-?,IL-10)in serum of rats in each group were detected by enzyme linked immunosorbent assay.Result:1.General situation: Before modeling,there was no significant difference in body weight,behavioral indicators and fecal properties of rats in each group(P > 0.05).After the establishment of the model,there was no obvious abnormality in the blank group and before the establishment of the model.The other rats in the model group had mental fatigue,dry and dark hair color,decreased food intake,weight growth,lean body,more stools,watery stools or shapeless stools.2.Abdominal wall reflex and diarrhea index score: compared with the blank group,the other model rats' abdominal wall reflex score decreased significantly(P < 0.001);compared with the blank group,the other model rats' diarrhea index score increased significantly(P < 0.001).It is suggested that the model can increase the visceral sensitivity of rats,and the rats have diarrhea symptoms.3.Open field experiment,tail suspension experiment and sugar water preference experiment:Compared with the blank group,the level score and vertical score of other open field experiment rats were significantly lower(P < 0.001);the tail suspension experiment score of other model rats was significantly higher(P < 0.001);the sugar water preference experiment score of other model group rats was significantly lower(P < 0.001).It was suggested that this method can make the rats appear the state of liver depression.4.Body weight growth and food consumption: Compared with the blank group,the weight growth of the other model rats was slow on the 7th and 14 th day of model building,with statistical significance(P < 0.05);the average daily food consumption of the other model rats within one week was significantly reduced(P < 0.05).It was suggested that this method can make rats appear spleen deficiency.5.Pathological changes of colon in rats: Compared with the blank group,there was noobvious congestion,edema,erosion and inflammatory cell infiltration in the colon of other rats.It was suggested that this method is consistent with the pathological changes of IBS-D.6.General situation: After treatment,compared with the model group,the rats in Shu-gan-jian-pi decoction group and Si-lian-kang group had better mental state,increased activity,gradually improved hair color and luster,loud voice,no obvious irritation,increased food intake,gradually formed faeces.7.Body weight change: After treatment,compared with the model control group,Shu-gan-jian-pi decoction group could significantly increase the body weight of rats(P <0.05),and there was no significant difference in the body weight growth of rats in Si-lian-kang group(P > 0.05).It was suggested that Shu-gan-jian-pi decoction can significantly improve the weight change of IBS-D rats.8.Abdominal wall rutting reflex: After treatment,compared with the model control group,there was no significant difference in the pain threshold of Si-lian-kang group(P > 0.05);Shu-gan-jian-pi decoction group could significantly increase the pain threshold of rats(P <0.05),suggesting that Shu-gan-jian-pi decoction could significantly reduce the visceral sensitivity of rats.9.Changes of inflammatory cytokines(IFN-?,IL-10)in serum of rats: After treatment,compared with the blank group,the level of IL-10 in serum of rats in model control group decreased significantly(P < 0.05),and the level of IFN-? increased significantly(P < 0.05).Compared with the model control group,the serum levels of IL-10 and IFN-? in Shu-gan-jian-pi decoction group and Si-lian-kang group increased significantly(P < 0.05)and decreased significantly(P < 0.05).Compared with the Si-lian-kang group,the serum level of IL-10 and IFN-? in Shu-gan-jian-pi decoction increased significantly(P < 0.05)and decreased significantly(P < 0.05).10.Changes of brain gut peptide(5-HT,VIP)in serum of rats: After treatment,compared with the blank group,the levels of 5-HT and VIP in serum of rats in model control group were significantly higher(P < 0.05).Compared with the model control group,the serum levels of5-HT and VIP in Shu-gan-jian-pi decoction group and Si-lian-kang group decreased(P <0.05).Compared with Si-lian-kang group,Shu-gan-jian-pi decoction group could significantly reduce the VIP level(P < 0.05),but there was no significant difference in 5-HT level(P >0.05).Conclusion:1.In this experiment,"chronic restraint stress + tail clip stimulation + senna leaf perfusion" was selected as the superposition compound model.By observing the general situation,AWR,diarrhea index,open field experiment,tail suspension experiment,sugar water preference,weight growth,food intake,colon HE staining and other comprehensive evaluation points of rats,the results were in line with the IBS-D animal model standard of liver depression and spleen deficiency,suggesting that the model was successful.1.Shu-gan-jian-pi decoction could improve the symptoms of IBS-D model rats with liver depression and spleen deficiency,increase the weight change of rats,increase the gut pain threshold of rats,and reduce the visceral sensitivity of rats.3.The mechanism of Shu-gan-jian-pi decoction in the treatment of diarrhea type irritable bowel syndrome may be related to the decrease of brain gut peptide such as 5-HT and VIP in serum.4.The mechanism of Shu-gan-jian-pi decoction in the treatment of diarrhea type irritable bowel syndrome may be related to the decrease of IFN-? and the increase of IL-10. |
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