| Objective: To observe and compare the effects of Liver Soothing and spleen strengthening components of Tongxieyaofang on visceral sensitivity,colonic pathology,expression of NGF and BDNF in colon and hippocampus,and changes of intestinal flora in IBS-D rats,aim to explore the mechanism of Tongxieyaofang Liver Soothing components Paeonia lactiflora and Fangfeng in treating visceral hypersensitivity of IBS-D rats on the basis of microbiota-gut-brain axis,so as to provide certain support for the clinical application of Tongxieyaofang and the treatment of IBS-D.Methods:40 male Wistar rats were randomly divided into control group(CON group),model group(Mo group),tangerine peel Atractylodes macrocephala group(C-B group)and Paeonia lactiflora Fangfeng group(B-F group)according to body weight,with 10 rats in each group.The IBS-D visceral hypersensitivity rat model was established by colorectal dilatation combined with chronic restraint stress for 14 days,and the rats in con group were stroked perineum daily for sham stimulation.After the completion of the model,the AWR score was used to detect the intestinal sensitivity of the rats in each group.The rats were fed in single cage,and the feces were collected to detect the water content of feces.Rats in each group were gavaged with 5ml / kg solution volume,rats in con group and Mo group were gavaged with distilled water,rats in C-B group were gavaged with 0.405g/ml tangerine peel and Atractylodes macrocephala extract solution,rats in B-F group were gavaged with 0.27g/ml Paeonia lactiflora extract solution for 14 days.After the treatment,the abdominal retraction reflex score and the water content of stool were measured again.The fecal samples of each group were collected to detect the intestinal flora.The colonic tissue was stained with hematoxylin red to observe the pathological changes;The protein expression of NGF and BDNF in colon and hippocampus was detected by ELISA.The structure of intestinal flora was observed by16 S r DNA technology.Results:1.General conditions: before modeling,the rats in each group had good spirit,healthy fur and normal stool;after modeling,the rats had poor spirit,lazy movement,disordered hair,weak stool and decreased food intake;after treatment,the mental state and hair health of the rats in the treatment group were improved,food intake recovered,and stool basically formed,and the overall state of the rats in the B-F group was better.2.Weight gain: after modeling,the weight gain of rats in each group was significantly lower than that in con group(P < 0.05);after treatment,compared with Mo group,the weight gain of rats in C-B group and B-F group were increased(P < 0.05,P < 0.05),and the weight gain of rats in B-F group was more than that in C-B group,but the difference was not significant(P > 0.05).3.Diarrhea(stool water content): after modeling,compared with con group,the stool water content of rats in each group increased significantly(P < 0.05);after treatment,compared with Mo group,the stool water content of rats in B-F group decreased significantly(P < 0.05),and the stool water content of rats in B-F group was lower than that of C-B group(P < 0.05).4.Visceral sensitivity evaluation: after modeling,compared with con group,visceral pain threshold of each group decreased significantly(P < 0.05);after treatment,compared with Mo group,visceral pain threshold of C-B group increased,but the difference was not significant(P > 0.05),visceral pain threshold of B-F group increased significantly(P <0.05);compared with C-B group,visceral pain threshold of B-F group was higher(P <0.05).5.Hematoxylin staining of colon: There was no organic lesion,which was consistent with IBS-D pathology.6.The expression of NGF and BDNF in hippocampus and colon: compared with con group,the expression of NGF and BDNF in hippocampus of Mo group was significantly decreased(P < 0.05,P < 0.05),and the expression of NGF and BDNF in colon was significantly increased(P < 0.05,P < 0.05);Compared with Mo group,the expression of NGF and BDNF in hippocampus of C-B group increased(P > 0.05,P > 0.05),and the expression of NGF and BDNF in colon decreased(P > 0.05,P > 0.05),but the difference was not statistically significant;In BF group,the expression of NGF and BDNF in hippocampus increased(P < 0.05,P <0.05),and the expression of NGF and BDNF in colon decreased(P < 0.05,P < 0.05).Compared with C-B group,the expression of BDNF in hippocampus and colon of B-F group was higher(P > 0.05)and lower(P > 0.05),but the difference was not obvious.The expression of NGF in hippocampus and colon was higher(P > 0.05)and lower(P< 0.05).7.Analysis of intestinal microflora structure: compared with con group,the abundance of lactobacilli and bifidobacteria in Mo group decreased(P < 0.05,P < 0.05),and the abundance of enterobacilli increased(P < 0.05);Compared with Mo group,the abundance of bifidobacteria increased and Enterobacteriaceae decreased in C-B group,but the difference was not significant(P > 0.05,P > 0.05),while lactobacilliaceae increased significantly(P < 0.05).Compared with Mo group,the abundance of lactobacilli and bifidobacteria in B-F group increased(P < 0.05,P < 0.05),and the abundance of Enterobacteriaceae decreased(P < 0.05).Conclusion:1.Radix Paeoniae Alba and Radix Saposhnikoviae can reduce visceral pain threshold of IBS-D rats.2.The changes of intestinal flora and the differential expression of NGF and BDNF in colon and hippocampus may be involved in the formation of visceral hypersensitivity of IBS-D.3.The Liver Soothing component of Tongxieyaofang,Radix Paeoniae Alba and Radix Saposhnikoviae,may treat IBS-D visceral hypersensitivity by improving the structure of intestinal flora and regulating the expression of NGF and BDNF,that is regulating the microbiota-gut-brain axis. |
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