Experimental Study On The Effect Mechanism Of IBS-C Rats In The Treatment Of Tongfujiangzhuo Decotion By The Correlation Between Neural Regulation And Intestinal Microorganism

Objective:This experiment through the observation on the effect of Tongfujiangzhuofang Decotion on rats with IBS,in the general state of rats,fecal water content and fecal grain number,body weight of rat,intestinal pathology,visceral sensitivity,intestinal mucosa 5-HT and 5-HTR3 expression,and the relationship of 5-HT and 5-HTR3 and Intestinal flora,explore the possible mechanism of treatment of IBS-C by the Tongfujiangzhuofang Decotion.Methods:To establish IBS-C rat model by using ice water irrigation method,randomly divide the 49 rats into 7 groups,The normal group（N）,model group（M）,western medicine group（X）,prebiotics group（Y）,chinese medicine high-dose group（ZH）,chinese medicine middle-dose group（ZM）,chinese medicine low-dose group（ZL）.After the model is established,the western medicine group took the xisha pelli tablet(3.6mg.kg^-1.d^-1),the prebiotics group took lactose oral solution(1667.5mg.kg^-1.d^-1),the Chinese traditional medicine group took Tongfujiangzhuo Decotion(18.5,9.25,4.625g^.kg^-1.d^-1),the model group was given an equal amount of saline irrigation.At the same time,the rats in each group were fed normally and observed the rat excrement in real time.After 14 days,and the fasting water 24h,each rat retained no less than 0.5g of feces,putting it in the frozen storage tube,and put in-80 refrigerator to save.To analyze the changes of fecal flora in rats by 16S rDNA amplification subsequencing technique.The rats were killed by the severed head,and the colon specimens were taken.To observe the pathological changes of intestinal tissues by HE staining,and the expression levels of 5-HT and 5-HTR3 in intestinal tissues by immunohistochemistry.Results:1.The general stateRats in normal group were in good condition with soft feces and yellow color.The rats in the mould group were prone to irritability,the hair was messy,the diet was reduced,the stool is hard,white and small.After treatment,fecal texture became soft,the quantity increased and the mood improved.2.Fecal water content and fecal grain numberAfter making the model,Compared with N,the fecal water content and grain number of each mould group decreased significantly（P<0.05）.There was no significant difference in fecal water content and fecal grain number between each mould group（P>0.05）,but compared with N,they were still less（P<0.05）.Compared between treatment groups,the fecal water content and fecal grain number of ZH,ZM,ZL,Y were more than X（P<0.05）.Compared between Z and X,the difference was not statistically significant（P>0.05）,The results of ZH were the closest to the normal group.3.The measurement of visceral sensitivity in ratsAfter making the model,The minimum capacity threshold of rectal balloon dilation was decreased in the model group（P<0.05）.After treatment,compared with M,The threshold of each treatment group increased（P<0.05）.Between the treatment groups,there was no significant difference between Chinese medicine groups（P>0.05）,significant differences remained between the remaining groups（P>0.05）.Compared with N,the Chinese medicine groups were closest to it,and had no significant difference（P>0.05）,the other groups were still significantly different（P<0.05）.4.The weight change in ratsThe weight of rats in each group increased steadily during the experiment,and the growth trend was basically consistent with no significant difference（P>0.05）.5.The pathological form of intestinalThe intestinal structures of rats were clearly visible in each group,and had no abnormal pathological changes.6.Immunohistochemical methods were used to detect the expression of5-HT and 5-HTR3 in intestinal tissues.The mean photometric values of 5-HT and 5-HTR3 were positively corre-lated with the staining depth of rats in each group.Compared with N,the 5-HT immunohistochemical staining positive cells in each treatment group and M were darker,the average Luminosity values is higher（P<0.05）.The 5-HTR3immunohistochemical staining positive cells in each treatment group and M were more shallow,the average luminosity values were lower（P<0.05）.Compared with M,5-HT staining in each treatment group was more shallow,the average Luminosity values is lower（P<0.05）,The 5-HTR3 staining in each treatment group was more darker,the average Luminosity values is higher（P<0.05）,and the change in ZH is more obvious.Between each treatment group,ZH and Y was more similar to each other（P>0.05）,and they were more closer to N.Between the other treatment groups,their differences were statistically significant（P<0.05）.7.Intestinal flora changes.The rarefaction curve flattens out,it shows that the sample size is sufficient.Compared with M,ACE index of N is more big,and the ACE index of each treatment group rise.ZH is the closest to Y.THE Shannon index between each group has no significant change.There were significant changes in the microflora structure of each group.Compared with N,acteroidetes of M are significantly higher,and firmicutes,fcidobacteria,verrucomicrobia of it are significantly reduced;The microflora structure of each treatment group is close to the normal group,and the effect of ZH is most obvious.8.Analysis of the correlation between neural regulation and intestinal microecology.5-HT is positively correlated with ACE and chao1 index,5-HTR3 is the opposite.5-HT and 5-HTR3 have a significant correlation with 35 species.They have no correlation with shannon and Simpson index.It is suggested that the expression of 5-HT and 5-HTR3 in the intestine interact with the richness of the bacteria,but have no interact with pathogen overgrowth.In a word,neural regulation has a significant correlation with intestinal microecology,Tongfujiangzhuo Decotion has the adjustment function to it.Conclusions:1.Tongfujiangzhuo Decotion can improve constipation,may by reducing the expression of 5-HT in rat intestinal tissue,and increasing the expression of5-HTR3,to improve intestinal sensitivity and abnormal powe,playing a role in the treatment of IBS-C.2.Tongfujiangzhuo Decotion can improve the intestinal microbial richn-ess in rats,adjust its microflora structure,stable intestinal microecology,This may be one of the mechanisms for its treatment of IBS-C.3.The correlation between neural regulation and intestinal microecology can affect the disease and treatment of IBS-C.