Effects Of Moxibustion On Gut Microbiome In Rats With Stomach Heat Syndrome Based On The Theory Of "Moxibustion With Heat Syndrome"

Objective:To observe the effects of moxibustion on body weight,gastric mucosal injury index,pathomorphology and intestinal microecology of rats with gastric mucosa injury in SH(Stomach Heat syndrome,SH)and SC(Stomach Cold syndrome,SC)by 16S rDNA sequencing technique,so as to explore the scientific nature of the theory of "Moxibustion with heat syndrome".Methods:48 male SD rats were randomly divided into 7 groups:The Control group,SH model group,SC model group,SH+ST(Stomach meridian acupoint,ST)group,SC+ST group,SH+GB(Gallbladder meridian acupoint,GB)group and SC+GB group.The gastric mucosal injury model of SH was established by intragastric administration of pepper suspension and anhydrous ethanol,and the gastric mucosal injury model of SC was established by intragastric administration of ice water at 4℃ and NaOH.After the modeling,the two syndrome diseases were intervention by moxibustion stomach meridian acupoints("Liangmen acupoint(ST21)" and "Zusanli acupoint(ST36)")and gallbladder meridian acupoints("Riyue acupoint(GB24)" and "Yanglingquan acupoint(GB34)")for 7 days.From the day before the beginning of the experiment,the weight of the rats was weighed every day to the end of the intervention,and the changes of the body weight of the rats were observed during the experimental period.After the intervention,the gastric mucosal injury index of all groups was observed and calculated,and the gastric mucosal tissue and fecal samples of all rats were collected.Gastric mucosal tissue was observed by HE staining under light microscope.The fecal samples were sequenced by 16s rDNA to detect the structural changes and relative abundance expression of intestinal microbiota in rat feces.Results:1.The results of body weight changes of rats:After the beginning of the model,the body weight of rats with SH syndrome and SC syndrome decreased gradually,and there was significant difference from the Control group on the 4th day(P<0.05).After moxibustion at stomach meridian acupoints,the body weight increased gradually.On the 10th day,the syndrome model group was significantly different from the gallbladder meridian acupoint group(P<0.05).2.Pathomorphological results:Gastric mucosal injury was obvious and inflammatory fine cells increased in SH group and SC group.Compared with the corresponding syndrome model group,the mucosal structure of moxibustion stomach meridian acupoint group was intact,the inflammatory cells’ number was reduced,and the pathology was significantly improved.There was no obvious recovery in the moxibustion gallbladder meridian acupoint group,which was basically the same as the corresponding syndrome group.3.Results of gastric mucosal injury index:Compared with the normal group,the gastric mucosal injury index of SH group and SC group increased significantly(P<0.05).Compared with the corresponding syndrome model group,the gastric mucosal injury index in the moxibustion stomach meridian acupoint group was significantly lower than that in the corresponding syndrome model group(P<0.05).The gastric mucosal injury index of moxibustion gallbladder meridian acupoint group decreased,and there was no statistical difference between moxibustion gallbladder meridian acupoint group and corresponding syndrome model group(P<0.05).4.The results of 16S rDNA sequencing of feces:After establishing gastric mucosal injury models of SH and SC respectively,the proportion of dominant bacteria at Phylum,Family and Genus level changed,and the composition of community and intestinal microbial structure were different.Through LEfSe analysis,compared with the Control group,the number of Bacteroides,Bacteroidia,Bacteroidales Rikenellaceae Ruminococcus gnavus and Porphyromonadaceae were significantly decreased in the SH group and SC group,and the decreased Ruminococcus and Porphyromonadaceae were significantly increased in the SH+ST group and SC+ST group respectively(P<0.05).The pathogenic bacteria,such as Collinsella,Coriobacteriales,Coriobacteriaceae,Coriobacteriia,Enter obacter i aceae and Enterobacteriales in SH group were significantly increased(P<0.05).In the SC group,the pathogenic bacteria such as Proteobacteria and Clostridium were significantly increased(P<0.05).Then,after the treated by moxibustion at stomach meridian points,the number of Collinsella,Coriobacteriales,Coriobacteriaceae and Coriobacteriia were significantly decreased(P<0.05).It also significantly increased the number of probiotics,such as Actinobacteria,Bifidobacteriales,Bifidobacteriaceae and Bifidobacterium(P<0.05).Moxibustion on the gallbladder meridian acupoint had fewer significant changes on the abundance of microflora.In the SH+GB group,there were more Blautia,producta and un_f_Alcaligenaceae(P<0.05),while in the SC+GB group,there were more Deltaproteobacteria,Desulfovibrionales and Desulfovibrionaceae(P<0.05).Conclusion:Moxibustion on gastric meridian acupoints has a significant effect on gastric mucosal injury in SH syndrome and SC syndrome,and can improve the weight loss,general behavioral changes and pathomorphological changes of gastric mucosa in rats with SH syndrome and SC syndrome.At the same time,It has a positive regulating effect on intestinal microecology,reducing the relative abundance of pathogenic bacteria and promoting the increase of probiotic abundance.As a result,the injury and repair of digestive system and gastric mucosa were improved.This paper reveals the acupoint specificity of moxibustion at stomach meridian acupoints in the treatment of gastric mucosal injury in SH syndrome and SC syndrome and its regulation mechanism on intestinal microecology,which shows that the theory of "Moxibustion with heat syndrome" is scientific and feasible.