| Objective1.On the basis of establishing the model of irinotecan tumor-bearing mice with delayed diarrhea,comparing the effects of traditional Chinese medicines Shengjiang Xiexin decoction and Huangqin decoction on delayed diarrhea caused by irinotecan.2.Combined with the bioinformatics analysis method of intestinal flora,to explore the effect of irinotecan chemotherapy on intestinal tract and the adjustment effect of the two formulas on intestinal tract after chemotherapy.Methods1.Establish irinotecan-induced delayed diarrhea tumor-bearing mice model by referring to the literature and preliminary experiment;2.Weight changes and diarrhea of mice in each group were observed during the experiment.Neutrophils were counted by rees-giemsa staining and evaluated in each group.Intestinal mucosal injury of mice in each group was evaluated by HE staining of intestinal tissue.Through the detection of the expressions of inflammatory factors and anti-inflammatory factors in colon tissues by Elisa method and real-time quantitative PCR technology,the alleviating effect of the two sides on the intestinal inflammatory response caused by irinotecan was explored.3.Colonic contents were collected and analyzed using illumina-miseq PE300 high-throughput sequencing platform for the microbial community and species composition of the samples in each group,including the analysis of a diversity,β diversity and species distribution,to verify the adjustment effect of Shengjiang Xiexin decoction and Huangqin decoction on intestinal flora after irinotecan chemotherapy.Results1.Through the evaluation of general situation,weight change and diarrhea score of model mice,it was confirmed that the model of irinotecan-induced diarrhea tumor-bearing mice was established successfully.2.After irinotecan intraperitoneal injection,the weight of mice in the model group gradually decreased,while that in the traditional Chinese medicine group began to rise on the 7th day,and there was a significant difference between the Shengjiang Xiexin decoction group and the model group from the 7th day to the 9th day(P<0.05),while there was a difference between the model group and the Huangqin decoction group on the 8th and 9th day(P<0.05).In terms of diarrhea,after 24 hours of irinotecan injection,all mice in the model group began to have soft and watery stools,with redness and swelling of the skin and mucous membrane around the anus,and the diarrhea symptoms gradually worsened.On the 5th day of the experiment,all the mice had moderate to severe diarrhea,with watery stools and partially watery stools,and no purulent and bloody stools.The feces of Shengjiang Xiexin decoction group and Huangqin decoction group were slightly moist and soft,some of the feces were not formed,and the anus was moist and slightly red and swollen.Diarrhea scores in the model group were all higher than those in other groups(P<0.05),while diarrhea scores in the Chinese medicine treatment group were lower than those in the model group(P<0.05).The number of neutrophils in the diarrhea model group was lower than that in the control group.The neutrophils in the Shengjiang Xiexin decoction group and Huangqin decoction group were 4.83±0.75 and 3.83±0.75 per view,respectively,which were higher than those in the model group.In terms of intestinal mucosa pathology,it was observed that the colonic mucosa of mice in the model group was swollen,with irregular glandular structure,and a large number of villi were shortened or lost,accompanied by inflammatory cell infiltration.The shape of intestinal villi was intact and the arrangement of mucosa was regular.According to the results of expression of TNF-a and IL-10,Shengjiang Xiexin decoction group and Huangqin decoction group can inhibit the intestinal mucosal inflammation by reducing the inflammatory factor expression and increase inflammation suppression factor expression.Moreover,the effect of Huangqin decoction group in reducing TNF-a was better than that of Shengjiang Xiexin decoction group(P<0.05),and the up-regulation of IL-10 in Shengjiang Xiexin decoction group was better than that in Huangqin decoction group,but the difference was not statistically significant(P=0.122).The results of PCR were significantly different between the model group and control group(P<0.01).TNF-α、TLR4 mRNA expression was significantly decreased in the Shengjiang Xiexin decoction group and the Huangqin decoction group compared with the model group,and the difference was significant(P<0.05).After irinotecan chemotherapy,the Alpha diversity index of the samples decreased,and that of the Shengjiang Xiexin decoction group decreased significantly(P<0.05).Compared with the control group,the Bacteroidales S24-7g in the other three groups decreased significantly(P<0.01),the level of Enterobacteriaceae in the model group increased significantly(P<0.05);compared with the model group,the content of Lactobacillaceae in Huangqin decoction group increased significantly(P<0.05),while the content of Porphyromonadaceae decreased significantly(P<0.05).The levels of Bacteroidaceae and Verrucomicrobiaceae in Shengjiang Xiexin decoction group increased significantly(P<0.05).Conclusion1.There was no significant difference between Shengjiang Xiexin decoction and Huangqin decoction in alleviating diarrhea caused by irinotecan,intestinal mucosal injury,weight loss and neutropenia.2.Shengjiang Xiexin decoction and Huangqin decoction can relieve the delayed diarrhea by adjusting the distribution of intestinal flora.Compared with the model group,Huangqin decoction can effectively improve the content of Lactobacillaceae;and the level of Bacteroidaceae in Shengjiang Xiexin decoction group increased significantly. |
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