Researching The Mechanism Of Tongfujiangzhuo Prescription For The Treatment Of IBS-C Based On The Regulation Effect Of Intestinal Microflorae On HPA Axis

Objective: The purpose of the study was to investigate the effect of Tongfujiangzhuo Prescription on constipation-predominant irritable bowel syndrome?IBS-C?model rats through the general condition,the number and water content of feces,visceral sensitivity,the content of serum corticotropin-releasing hormone?CRH?,adrenocorticotrophic hormone?ACTH?,corticosterone?CORT?and intestinal microflorae,and then,try to further explore the possible mechanism of the Tongfujiangzhuo Prescription.Methods: 49 six-week-old male SD rats were randomly divided into seven groups,each group 7 rats.They were respectively marked for the normal group?N?,model group?M?,the cisapride group?X?,the prebiotics group?Y?,high dose group?ZH?,moderate dose group?ZM?,and low dose group of Tongfujiangzhuo Prescription?ZL?.Except the N group,the rest groups were controlled in order to make IBS-C animal models by ice saline.After the IBS-C model was established successfully,the N and M group were given normal saline,the other treatment groups received liquid gavage once a day.During the treatment,all groups had normal diet and drinking water.After 2weeks continuous treatment,observate the activity,fecal grains and water content of all rats.The visceral sensitivity of rats was determined by the method of colorectal balloon dilatation.Detected the content of serum CRH,ACTH,CORT by radioimmunoassay and the structure and quantity of intestinal microflora of rats in each group were detected with 16 Sr DNA amplification subsequencing technique.Results:1.The general state Compared with N group,the rats appeared fur lackluster,listless,irritableand overreact.The stool is bulbous,dry and hard,with grayish brown or grayish color.At the end of the treatment,The rats in each treatment group had different levels of improvements.General condition of rats in the ZH,ZM,ZL and Y groups were close to the N group.While the condition of X were slightly lower than the N group and the recovery of the M group was the worst.2.The comparison results of fecal grains and water content in each group Before the treatment,the N group compared with the other groups,the differences were statistically significant?P<0.05?.After the treatment,M group compared with each treatment group,statistically significant difference?P<0.05?.3.The comparison results of visceral sensitivity in each group Before the treatment,the N group compared with the other groups,the differences were statistically significant?P<0.05?.After the treatment,M group compared with N group,statistically significant difference?P<0.05?.Compared with M group,the difference of each treatment group was statistically significant?P<0.05?.4.The test results of serum CRH in each group Compared with the N group,the ZH group,no statistically significant difference?P>0.05?and the other groups,statistically significant difference?P<0.05?.There was statistically significant difference between the M group and each treatment group?P<0.05?.5.The test results of serum ACTH in each group Compared with the N group,the M group,no statistically significant difference?P>0.05?,while the X and M group,statistically significant difference?P<0.05?.There was statistically significant difference between the M group and each treatment group?P<0.05?.6.The test results of serum CORT in each group M group compared with N group,statistically significant difference?P<0.05?.There was statistically significant difference between the M group and each treatment group?P<0.05?.7.The results of microbial community structure changes in each group Compared with the N group,the intestinal microbial community abundance in the M group was significantly decreased,statistically significant difference?P<0.05?.The differences in intestinal microbial community composition among N and M group were the greatest.Compared with the N group,there are three phyla,seven families and four genera,such as firmicutes,lactobacillaceae and lactobacillus were significantly reduced,while bacteroidetes,four families and sixteen genera,such as prevotellaceae,prevotellaceae_NK3B31_group were significantly increased,statistically significant difference?P<0.05?.8.The regulation effect of intestinal microorganism on HPA axis The bacteroidetes,prevotellaceae,bacteroidaceae,porphyromonadaceae,bacteroides,alloprevotella,prevotellaceae_NK3B31_group and prevotella₉were positively correlated with CRH and ACTH.Porphyromonadaceae was positively correlated with CORT.Firmicutes was negatively correlated with ACTH.Acidobacteria and verrucomicrobia were negatively correlated with CRH and ACTH.Lactobacillaceae,bifidobacteriaceae,lactobacillus and bifidobacterium were negatively correlated with ACTH and CORT,statistically significant difference?P<0.05?.Conclusions:1.Tongfujiangzhuo Prescription,not only can improve the IBS-C rats general situation,increase the fecal grains and fecal water content,but also reduce the visceral sensitivity of IBS-C rats.2.The mechanism of Tongfujiangzhuo Prescription on the treatment of IBS-C may be by regulating intestinal flora to decrease the content of CRH,ACTH and CORT in serum,which can regulating the dysfunction of HPA axis and improve the state of intestinal hypersensitivity.