Mechanism Of Moxibustion In The Treatment Of Chemotherapyinduced Intestinal Mucositis Rats From The View Of Regulating Intestinal Microflora And NLRP3 Inflammasome

AIM The purpose of this study is to clarify the healing effect of moxibustion on the structure and function of intestinal mucosa injury through analyze the intervention of different therapeutic method on the intestinal mucoiditis rats.And analyze the mechanism from the point of intestinal microflora and immune factors,so as to provide a certain experimental basis and scientific basis for the clinical moxibustion curative effect of the treatment of IM.METHODS Rats from the moxibustion group were fixed on the moxibustion platform and the moxa stick was placed above the acupoints,1.5-3cm from the skin.Guanyuan(CV4),Qihai(CV6),Shenque(CV8),Zhongwan(CV8)and Tianshu(ST25,bilateral)were treated with mild moxibustion for 15 min once a day for fifteen days.The rat model of IM was established by intraperitoneal injection of 5-Fu oneoff.The general status of rats were recorded daily when the modeling began.Hematoxylin and eosin staining;Electron microscopy;Enzyme-linked immunosorbent assay;Western blotting;TUNEL technology;q PCR;Immunofluorescence;16S r RNA amplicon sequencing.RESULTS The general score of the model group was significantly higher than that of the control group after modeling(P<0.01),and the body weight and foodintake of the model group were significantly lower than those of the control group(P<0.01 or P<0.01),and the diarrhea rate from the model group was higher than that of the control group.Pathological changes of intestinal mucosa in the model group were observed.ELISA showed that the level of serum TNF-? in the model group was significantly higher than that in the control group(P<0.01),on the contrary,the level of IL-10 was significantly lower(P<0.01).It is suggested that the rat model of intestinal mucoiditis induced by 5-Fu was established successfully.The scores of general condition in the three groups were significantly higher than those in the control group(P<0.01).From the 3rd day after chemotherapy,the body weight or foodintake in the model group were lower than those in the moxibustion group and probiotics group,and the diarrhea rate was higher than that in the moxibustion group and probiotics group.The pathological examination showed that the degree of intestinal mucosa injury in the model group was significantly higher than that in the control group and moxibustion group(P<0.05 or P<0.01),and the expression of tight junction protein Occludin and ZO-1 was significantly higher than that in the model group after moxibustion or probiotics treatment(P<0.05).Under microscope,the apoptotic bodies increased in model group,but few apoptotic cells were found in moxibustion group and probiotics group.Both immunofluorescence and q PCR showed that the expression of NLRP3 in the model group was significantly increased,and moxibustion or probiotics could decrease the expression of NLRP3.The serum level of TNF-?,IL-6,IL-1? and IL-18 in the model group were significantly higher than those in the control group(P<0.01),moxibustion group and probiotics group(P<0.05 or P<0.01).The expression of NLRP3,Caspase-1,Gasdermin D relative protein expression in the model group was significantly higher than that in the control group,moxibustion group and probiotics group(P<0.05 or P<0.01).The Shannon index of rats from moxibustion group was significantly higher than that of model group(P<0.05);The results of PCo A analysis showed that the similarity of intestinal microflora structure between moxibustion group and probiotics group was higher than that of model group.Compared with control group(P<0.01),moxibustion group(P<0.05 or P<0.01)and probiotics group(P<0.05 or P<0.01),in the model group the proportion of Lactobacillus was decreased significantly,and the proportion of Roseburia and Escherichia was significantly increased,which are all colitis-associated bacteria.There was a significant correlation between the three kinds of bacteria and inflammatory factors such as NLRP3,IL-1?,IL-18,IL-6 and TNF-?(P<0.05 or P<0.01).CONCLUSION The intestinal mucositis model induced by one-time intraperitoneal injection of 5-Fu was simple in operation,and had short cycle and good stability.Mild moxibustion can alleviate the degree of intestinal mucosal injury,increase intestinal probiotic bacteria,inhibit harmful bacteria and regulate the expression of NLRP3,Caspase-1 and related inflammatory factors,and thus treat IM caused by chemotherapy.The mechanism may be to regulate intestinal microflora homeostasis,and thus reduce the pyroptosis,so as to alleviate the inflammatory injury.