From The Regulation Of Intestinal Motility Based On Intestinal Flora To Explore The Effect Mechanism Of Yishen Qingli Heluo Prescription In The Treatment Of CKD

Objective:Established the 5/6 nephrectomy rat model to observe the effect of Yishen Qingli Heluo Prescription on renal function of 5/6 nephrectomy rats.Compared the effect of Yishen Qingli Heluo Prescription on intestinal motility of rats with 5/6 nephrectomy through a series of intestinal motility experiments.Found the dominant flora by analyzing the difference of intestinal flora of 5/6 nephrectomy rats in different groups.Explored the relationship between intestinal microecology,gastrointestinal symptoms and intestinal motility in CKD.Analyzed the influence mechanism of Yishen Qingli Heluo Prescription on intestinal flora to speculate the possible mechanism of Yishen Qingli Heluo Prescription on improving renal function of CKD and the relationship between intestinal motility and intestinal microecology of CKD.Method:24 adult SPF-grade male SD rats were randomly divided into 3 groups(n=8),sham operated group,model group,Yishen Qingli Heluo Granule(YQHG)group.2 weeks after the established of 5/6 nephrectomy rat model,the sham group and model group were gastrically filled with 0.9%sterile water daily,and the YQHG group was gastrically filled with YQHG daily for a total of 8 weeks.Observed the general condition,weight,the conditions of drinking and eating,and feces of the rats.Changes of gastrointestinal motility were measured by colorectal beading experiment,first defecation experiment,gastric emptying and small intestine propulsion rate experiment.24h urine protein quantification,Scr and BUN of rats were detected to assess rat renal function.Enzyme linked immunosorbent assay(ELISA)was used to detect 5-HT,ACh,spermine in serum and colon of rats.16S rRNA high-throughput sequencing was used to detect and analyze the intestinal flora of rats.Outcome:1.General situation of rats:After the surgery,the rats in the model group gradually appeared to be mentally weak,the coat color darkened,the rat hair was disheveled,the teeth turned yellow.The rats in the YQHG group appeared the similar condition like the model group,but it was better.Compared with the Sham group of rats,the model group gained more body weight while those in the YQHG group increased slowly.The water consumption of model group and YQHG group was higher than that of sham group,but the water consumption of model group increased significantly(P<0.05).The fecal water content of rats in the model group was lower than that in the sham group,and the fecal water content in the YQHG group was higher than that in the model group(P<0.05).There were differences in the 24h feces weight of the rats in each group,and the defecation volume in the YQHG group was significantly higher than that in the model group(P<0.05)2.Renal function:Scr,BUN and 24h urine protein of rats in the YQHG group were significantly reduced compared with the model group(P<0.01).Pathological changes such as glomerular and tubular lesions and inflammatory cell infiltration in the kidneys of rats in the YQHG group were improved compared with those in the model group.3.Intestinal motility:Compared with the sham group,the model group used a longer time to finish first defecation while the YQHG group used a shorter time.(P<0.001).Compared with the sham group,the colorectal propulsion of rats in the model group was slower,while the YQHG group had a significant advantage(P<0.05).Gastric emptying rates were significantly different in each group(P>0.05).Compared with the sham group,the small intestine advancement rate in the model group was lower and that in the YQHG group was higher.Compared with the sham group,the levels of serum serotonin(5-HT),colonic 5-HT,serum acetylcholine(Ach),colonic Ach and serum spermine in the model group were decreased;Serotonin(5-HT),colonic 5-HT,serum acetylcholine(Ach),colonic Ach and serum spermine of rats all increased(P<0.01).4.Gut microbiota:The gut microbiota was analyzed at the phylum and genus levels.Compared with the sham group,the intestinal flora of the model group was different significantly.The YQHG group had more similar intestinal flora composition to the sham group than the model group.In particular,the relative abundance of intestinal bacteria and community heatmap analysis at the phylum level showed that there were obvious differences between the intestinal flora of rats in the model group,such as Firmicutes and Bacteroidota.At the level of the phylum and genus,there were differences in the dissimilarity within the three groups.At the level of phylum and genus,the within-group difference of the model group was smaller than that of the sham group.At the phylum level,compared with the model group and sham group,the YQHG group has greater intra-group dissimilarity.At the genus level,the YQHG group has greater intra-group dissimilarity compared with the model group.Compared with the sham group,the within-group differences were small.At the phylum and genus levels,5/6 nephrectomy rats had altered gut flora diversity,while YQHG increased bacterial diversity at the phylum and genus levels.Conclusion:1.Yishen Qingli Heluo Prescription can reduce Scr and BUN in serum and 24-hour urinary protein of 5/6 nephrectomy rats,improve glomerular and renal tubular lesions to delay the progression of CKD.2.Yishen Qingli Heluo Prescription can promote ENS to release 5-HT and ACh,increase defecation volume,increase fecal water content,and promote intestinal motility of rats.3.Yishen Qingli Heluo Prescription can increase the diversity of intestinal flora and the abundance of beneficial bacteria in 5/6 nephrectomy rats,improve the intestinal microecology,and take the return of intestinal flora in 5/6 nephrectomy rats to normal.