The Effect Of Abdominal Massage On The Intestinal Microbiota In Immature Rats With Spleen Deficiency Diarrhea

ObjectiveThe effects of abdominal massage on the general condition,body weight,colonic histomorphology and intestinal flora of immature rats with spleen deficiency diarrhea were observed by using the mastication method.At the same time,the intestinal flora were examined by 16 S r DNA amplicon sequencing technique to explore the possible mechanism of abdominal massage on relieving the symptoms of spleen deficiency diarrhea in immature rats.This study is expected to provide a scientific and theoretical basis for clinical nurses to apply abdominal massage in the care of children with spleen deficiency diarrhoea.Methods3-week SPF-grade SD rats were experimentally treated.Grouping was carried out via the Excel random function Rand.Six rats were randomly selected as the normal group to continue basal feeding The remaining 18 rats were gavaged with a mixture of "Senna &Rhubarb"(3g/ml),1ml/100 g each,once daily for 7 days,and assessed for modeling results.After modeling the rats were randomly divided into three groups: the model group,CW-massage group and the CCW-massage group.The CW-massage group was given clockwise abdominal massage and the CCW-massage group was given counter-clockwise abdominal massage.The frequency of abdominal massage was 120-160 times/min,10 minutes each time,once a day for 14 days.During this period,the general condition of the pups in each group(including their mental activity,hair color,etc.),faecal properties and weight gain were observed and recorded daily.At the end of the experiment,the histopathological changes in the colonic mucosa were observed by HE staining.The changes in the structure and diversity of the intestinal flora of the colonic contents were detected by 16 S r DNA amplicon sequencing.Results1.The abdominal massage can improve the general condition,body weight,partially repair the damage to the intestinal mucosa of immature rats with spleen deficiency diarrhea.(1)General condition and faecal properties: After the abdominal massage intervention,the faces of the pups in both groups gradually changed from loose stools to soft stools to normal stools(hard and granular in texture).The pups in both groups showed a gradual increase in activity,and their hair color gradually changed from yellow and disheveled to soft and shiny;the arching of the back and curling up in piles were reduced;and the degree of perianal soiling was reduced.(2)Body weight: Body weight and its growth rate were increased in both mosquito groups after treatment intervention compared to the model group,but there was no statistical difference between the groups(P>0.05).(3)Colonic mucosal condition: After the mastication intervention,the two mastication groups partially reversed the condition of the model control group.In both groups,the villous structure was relatively intact,and the epithelial cells of the surface layer were slightly necrotic and detached;the submucosa was still slightly edematous and congested,but no significant difference was observed between the two groups.2.Mastication regulates the dysbiosis of intestinal flora caused by spleen deficiency diarrhea.(1)Beta diversity analysis: the intestinal flora community structure in the model oup was more different from that in the control group.The difference in community structure between the two massage groups and the control group was effectively reduced after mastication(R=0.646,P=0.001).(2)Analysis of differences in species abundance: in the model group,Akkermansia,Clostridium＿innocuum＿group,Ruminococcus＿torques＿group,Parabacteroides,Coriobacteriaceae＿UCG-002 significantly increased in abundance compared to the control group.After massage,Bifidobacterium,Lachnospiraceae＿NK4A136＿group,Blautia,and other genera were significantly increased in abundance compared to the model control and returned to near normal levels compared to the control;while Clostridium＿innocuum＿group,Ruminococcus＿torques＿group,Parabacteroides,Coriobacteriaceae＿UCG-002 were significantly decreased in abundance compared with the model control.(3)LEf Se analysis: Those enriched in the control group included(P < 0.01): g＿＿Dorea(LDA=4.45),g＿＿Bifidobacterium(LDA=4.51),g＿＿Faecalibaculum(LDA= 4.71),g＿＿Turicibacter(LDA=5.01),etc.Enriched in the model group included(P < 0.01):p＿＿Desulfobacterota(LDA=4.35)p＿＿Verrucomicrobiota(LDA=5.52)g＿＿Ruminococcus＿torques＿group(LDA=4.13),g＿＿Akkermansia(LDA=5.50),g＿＿Coriobacteriaceae＿UCG＿002(LDA=4.57),g＿＿Clostridium＿innocuum＿group(LDA=4.30),etc.After the CW-massage,the differential flora enrichedincluded(P < 0.01):p＿＿Firmicutes(LDA=5.31),g＿＿Cutibacterium(LDA=4.54),g＿＿Dorea(LDA=4.58),g＿＿Family＿XIII＿AD3011＿group(LDA=4.58)group(LDA=4.53),etc.After the CCW-massage,the differential flora enriched included(P < 0.01): p＿＿Actinobacteriota(LDA=4.77),o＿Bifidobacteriales(LDA=4.72),g＿＿DNF00809(LDA=3.97),g＿＿Defluviitaleaceae＿UCG＿011(LDA=3.80),etc.ConclusionsThe experimental results of this study suggest that the intestinal flora characteristics of immature rats with spleen deficiency diarrhea are associated with an increased abundance of harmful bacteria that disrupt the intestinal barrier,and perhaps not with inflammatory-related bacterial infections.The experimental results also demonstrated that the effect of mastication on young rats with deficient spleen diarrhea was manifested in three ways:(1)abdominal massage could alleviate the symptoms of deficient spleen diarrhea in immature rats.(2)The abdominal massage can repair some of the damage to the intestinal mucosa.(3)By reducing the abundance of bacteria that have harmful effects on the intestinal mucosal barrier and increasing the abundance of many types of beneficial bacteria,the mastication can regulate the disordered intestinal flora structure of immature rats with spleen deficiency relieve the symptoms of spleen deficiency diarrhea.This may be one of the mechanisms of the abdominal massage to relieve diarrhea in children with spleen deficiency.Finally,it was found that the difference between the two different directions of clockwise and counterclockwise abdominal massage on the intestinal flora,spleen deficiency symptoms,body weight and colonic mucosa of the immature rats was not significant.This study confirms that the abdominal massage is an effective technique of TCM for the treatment of paediatric diarrhoea and provides more scientific basis for its clinical practice and application.The efficacy of the abdominal massage is clear,it is safer than other therapies,and it is well accepted by children and parents.So it is recommended that the abdominal massage be used more often in clinical practice for children with spleen deficiency diarrhea.Nurses can also guide parents to learn abdominal massage and apply it consistently at home to increase the child’s compliance and continuity of treatment and to promote recovery from the disease.