| Splenasthenic diarrhea is a systemic pathological condition affecting many organs.It is veterinary clinical common disease,frequently-occurring diseases.In modern medicine,the diarrhea syndrome of spleen deficiency mostly belongs to the category of chronic and chronic diarrhea,so that loose matter,diarrhea,physical decline,loss of appetite,listlessness and inactivity are the main characteristics.The cause of the disease is complex,the course of the disease is difficult to control,and all seasons can come on,the health development of livestock and poultry breeding industry has a greater impact.There are no specific drugs for this disease,and antibiotics are often used in clinical treatment.Irregular use or abuse of antibiotics often occurs,which aggravates the frequent occurrence of diarrhea in animals,while drug resistance becomes more and more serious,bringing great harm to animal health.Therefore,clinical veterinary workers advocates the application of Chinese herbal medicine preparation to the prevention and treatment of the disease,"Lingzhu powder" is composed of poria,stir-baked Rhizoma Atractylodis macrocephalae with soil,Codonopsis Radix,Bran-processed Rhizoma Atractylodis,Citri Reticulatae pericarpium,Alismatis Rhizoma(stir-bake with saltwater),Zingiberis Rhizoma praeparatum.tonic temper,the effect of moisture penetration of benefit water.early clinical laboratory experiments confirmed that the compound can be effective prevention and control of animal Splenasthenic diarrhea.In order to explore the mechanism of its action,this study took rats as experimental animals to investigate the mechanism of "Lingzhu powder" intervention in the diarrhea of spleen deficiency in rats from the perspective of intestinal mucosal barrier.First,in order to determine the optimal therapeutic dose of Lingzhu powder in the intervention treatment of splenasthenic diarrhea in rats,80 rats were randomly divided into a blank control group(10)and a modeling group(70).The modeling group was randomly divided into model group,Lingzhu powder group A(40 g/kg·body weight),Lingzhu powder group B(20g/kg·body weight),Lingzhu powder group C(10 g/kg·body weight),Lingzhu powder group d(5g/kg·body weight),Lingzhu powder group E(2.5 g/kg·body weight)and Shenlingbaizhu powder group(20 g/kg·body weight)The clinical manifestations of all the experimental rats were observed and scored macroscopically before,after and after 14 days of drug intervention,respectively.The results showed that the macroscopical scores of the rats in the post-modeling group were significantly higher than those in the blank control group(P < 0.05).After the intervention of Lingzhu powder,the macroscopic integral of each group of rats decreased significantly(P < 0.05),and there was no significant difference between Lingzhu powder B group and blank control group(P > 0.05).The results showed that ling shu 20 g/kg·body weight dose intervention was the best.Second,in order to further explore the mechanism of "Lingzhu powder" in the intervention of splenasthenic diarrhea,40 rats were randomly divided into blank control group,model group,Lingzhu powder group and shenlingbaizhu powder group.In addition to the blank control group,the rats in the other three groups all adopted the method of "broken qi + under bitter conditions +Eating disorders" to establish the diarrhea syndrome model of spleen deficiency.After successful modeling,the rats in the Lingzhu powder group and the shenling baizhu powder group received gavage intervention treatment according to 20 g/kg·body weight,respectively,and blood and tissue samples of each group were collected on the 14 day after the intervention treatment.Thymus and spleen indexes of rats in each group were calculated,serum d-xylitose content of rats was determined by phloroglucinol method,serum DAO and D-LA levels were detected by enzyme-linked immunoassay,and routine paraffin sections of colon were prepared to observe the changes of colonic mucosal tissue structure.The results showed that,compared with the blank control group,the thymus index,spleen index and serum d-xylose content in the model group were significantly reduced(P < 0.05),serum DAO and D-LA levels were significantly increased(P < 0.05),and the paraffin section showed that the upper layer of the colon mucosa was detached and the integrity was damaged.After lingzhu powder intervention,the thymus and spleen index and serum d-xylitose content were significantly increased(P < 0.05),the serum DAO and d-la levels were significantly decreased(P < 0.05),the colonic mucosal epithelium was regenerated,and the integrity was restored.The results showed that "Lingzhu powder" could improve the intestinal absorption function of rats with spleen deficiency and diarrhea,and improve the intestinal damage caused by spleen deficiency and diarrhea.Third,to further explore the effect of Lingzhu powder on colonic mucosal immunity in rats with splenasthenic diarrhea.On the basis of test 2 design,2 cm of proximal colon was collected from rats in model group,blank control group,Lingzhu powder group and Shenlingbaizhu powder group,respectively.Intestinal mucosa was collected and homogenized.Supernatant was taken after centrifugation.The results showed that compared with the blank control group,the colonic mucosa levels of SIg A and IL-4 in the model group were significantly reduced(P < 0.05),while the levels of TNF-α and IFN-γ were significantly increased(P < 0.05).After Lingzhu powder intervention,the levels of SIg A and IL-4 in the colonic mucosa of rats were significantly increased(P < 0.05),while the levels of TNF-α and IFN-γ were significantly decreased(P < 0.05).The results showed that Lingzhu powder could modulate the immune function of colonic mucosa in rats with splenasthenic diarrhea.Fourth,to further investigate the effect of Lingzhu powder on the expression of Claudin-1,Occludin and ZO-1 mrna of tight junction proteins in the colon of rats with splenasthenic diarrhea.On the basis of experimental design,the proximal colon of rats in model group,blank control group,Lingzhu powder group and Shenlingbaizhu powder group was extracted about 2 cm,Trizol method was used to extract total RNA,and Quantitative real-time PCR method was used to detect the expressions of Claudin-1,Occludin and ZO-1 mrna in the colon of rats with spleen deficiency and diarrhea.The results showed that the expressions of claudin-1,occludin and zo-1mrna in the model group were significantly lower than those in the normal control group(P <0.05),and the expressions of Claudin-1,Occludin and ZO-1mrna in the model group were significantly higher after the intervention of Lingzhu powder group(P < 0.05).The results showed that Lingzhu powder could repair the tight connection structure and integrity of colonic epithelial cells in rats with splenasthenic diarrhea,and protect the barrier function of intestinal mucosa.In summary,Lingzhu powder can reduce the macroscopic integration of splenasthenic diarrhea,improve the content of serum d-xylose,and restore the gastrointestinal absorption function.The indexes of spleen and thymus were increased,the levels of TNF-α and IFN-γ in the colonic mucosa of rats with spleen deficiency and diarrhea were decreased,and the levels of IL-4and SIg A in the colonic mucosa were increased,so as to play the role of immune regulation.It can effectively improve the repair of colonic mucosal tissue injury in rats with splenasthenic diarrhea,reduce the content of D-LA in serum and the activity of DAO in rats with diarrhea syndrome of spleen deficiency,up-regulate the expression levels of intestinal mucosa zo-1,claudin-1 and occludin m RNA in rats with splenasthenic diarrhea,thereby reducing the permeability of intestinal mucosa and protecting the intestinal mucosal barrier. |
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