Effect And Mechanism Of Moxibustion In Regulating Gastrointestinal Motility In Functional Gastrointestinal Diseases

【Objectives】Moxibustion is an effective TCM nursing technique for functional gastrointestinal diseases(FGIDs),and its pathogenesis suggests it is very suitable for nursing research.However,moxibustion treatment of FGIDs has a variety of ways,and the mechanism of action is not yet clear,which has become an obstacle to the promotion and application of moxibustion.In this paper,animal comparative experiment was designed to explore the best moxibustion method according to the possible influence of related factors on the effect of moxibustion method commonly used in the treatment of the disease.Then using modern biological technology,based on hyperpolarization activated cyclic nucleotide gated cation channel(HCN)experiment to study the effect and mechanism of moxibustion in the treatment of FGIDs.【Methods】1.Literature researchThe application of traditional Chinese medicine nursing techniques in digestive tract diseases,moxibustion treatment of FGIDs,HCN and packing were studied in literature,and the optimal moxibustion treatment of FGIDs was compared.The possible pathway of moxibustion treatment of FGIDs was analyzed and the mechanism of action was discussed.To analyze the feasibility of FGIDs rat model making,moxibustion,HCN ion channel and FGIDs related experimental techniques.Construct research framework and draw detailed technical roadmap.2.Moxibustion researchSPF SD rats were divided into normal group,model group,Domperidone positive control group,moxibustion group,ginger moxibustion group,moxa fumigation group and hot moxibustion group.Combined stress intervention(Guo’s tail-clipping stimulation + hunger and satiety disorder)was used to stimulate the rats for 4 weeks to establish FGIDs rat model.Mice in normal group,model group and positive control group: head,tail and limbs of mice were fixed with fixators without any treatment.The fixators were grabbed and fixed for 20 min every day.Positive control group was orally administrated with Domperidone and model group was administrated with the same amount of normal saline.Mice in the moxibustion group: the head,tail and limbs of mice were fixed with fixators,and the mice were treated with RN12(Zhongwan)and ST36(Zusanli)moxibustion with moxa sticks for 20 minutes every day,and the same amount of normal saline was given by gavage.In the ginger moxibustion group,the head,tail and limbs of mice were fixed with a fixator.RN12 and ST36 were treated with ginger moxibustion for 20 min.The positioning was the same as above,and the same amount of normal saline was given by gavage.In the moxa fumigation group,the rats’ head,tail and limbs were fixed with fixers,and the amount of moxa sticks used in the moxibustion group was fumigated for 20 min every day,and the same amount of normal saline was given orally.In the hot moxibustion group,the head,tail and limbs of mice were fixed with a fixator,and the mice were treated with hot moxibustion RN12 and ST36 for 20 min every day,followed by the same amount of normal saline.The above treatment is carried out every day(7:00-9:00)for 6 consecutive days,1 day rest,7 days as a course of treatment,for 2courses of treatment.The general survival status of mice in each group was observed.The food intake,body weight and sugar and water consumption of rats in each group were detected.The gastric residual rate and small intestinal propulsion ratio of mice in each group were measured.3.Effect and mechanism of HCN moxibustion in the treatment of FGIDs.SPF SD rats(200±20g,6 to 8 weeks of age)were randomly divided into normal group,model group,Domperidone positive control group,moxibustion group and HCN blocking group.Model making and processing are the same as above.In addition,the HCN blocking group was injected with ZD7288 blocker,and the model group,morpholine positive control group,and moxibustion group were injected with the same amount of normal saline.In addition to the above experimental indicators,the protein expression of HCN1 and HCN2 in gastric tissue(gastric antrum above the greater curvature of the stomach)was detected by WB,the expression of HCN1,HCN2 in the upper small intestine and medial hypothalamus nucleus of rats in each group was detected by QPCR,and the c-kit and HCN1 in gastrointestinal parts of each group were detected by double immunofluorescence labeling method.【Results】1.Literature researchTraditional Chinese Medicine nursing technology moxibustion for FGIDs has definite effect on intervention.Moxibustion and separation moxibustion are the most common.The chemical effect of moxa smoke and the radiation effect of Moxa fever play a major role in moxibustion.GI motility disorder is the main factor of FGIDs.Moxibustion therapy can regulate GI motility.HCN ion channels were expressed in the hypothalamus,gastric antrum and upper small intestine.HCN ion channels are related to autonomic pacing,and HCN is coexpressed with GI slow-wave pacing cells ICCs.The methods of making rat models for FGIDs experiments were mature,and the techniques of moxibustion,HCN ion channels and FGIDs related experiments were mature.Complete the research framework construction and technical roadmap drawing.2.Moxibustion researchCompared with the model group,the moxibustion group,moxa fumigation group hot moxibustion group and ginger moxibustion group all improved.Compared with moxa fumigation group,hot moxibustion group and ginger moxibustion group,moxibustion group had the best improvement effect on food intake,weight,sugar and water consumption,gastric residual rate and small intestine propulsion ratio of experimental rats.Therefore,in this animal experiment,moxibustion is the best moxibustion method for FGIDs.At the same time,the experiment also found that the moxibustion group and the positive group of rats have a great difference in each index,and with the extension of intervention time increased trend.3.Effect and mechanism of HCN moxibustion in the treatment of FGIDs.Compared with the model group,the experimental rats in the moxibustion group were improved,and all the observed measurement indexes were statistically significant(P<0.01).However,there were significant differences in various indicators between the moxibustion group and the blocking group,suggesting that the treatment of functional gastrointestinal diseases in rats by blocking HCN was ineffective.In conclusion,moxibustion is likely to regulate GI motility in FGIDs rats through HCN.Study found that moxibustion group and positive group compared with model group rats,the experimental rats after intervention,WB measurement stomach big curved upper HCN1,engineered ion channel protein expression in gastric antrum tissue increase,significant statistical significance(P > 0.01),QPCR detection on the small intestine and the medial nuclear HCN1,engineered ion channel expression was also significantly increased,significant statistical significance(P > 0.01),but still have large differences between moxibustion group and positive group,with statistical significance(P < 0.05).It is suggested that both moxibustion and Domperidone can improve the expressions of HCN1 and HCN2 ion channels in the detected tissues of rats with FGIDs,but there is a great difference in the intensity of action between them.Compared with the HCN blocking group,the expressions of local HCN1 and HCN2 ion channels in the moxibustion group were significantly different,suggesting that moxibustion lost the regulation effect of HCN1 and HCN2 ion channel expression in the detected tissue of FGIDs rats after blocking HCN ion channels.The expressions of c-kit and HCN1,c-kit and HCN2 in the gastric antrum of upper curvature of stomach and the intestinal tissue of upper small intestine were observed by double immunofluorescence labeling in each group.It was found that in the model group,the red fluorescein-labeled c-kit and green fluorescein-labeled HCN1 and HCN2 ion channels were significantly less than those in the normal group.In the moxibustion group and the positive group,the red fluorescent-labeled c-kit and green fluorescent-labeled HCN1 and HCN2 ion channels were significantly more than the model group.In the moxibustion group,the green fluorescently labeled HCN1 ion channel was evenly distributed and accompanied with the red fluorescently labeled c-kit,that is,the HCN1 ion channel was co-expressed with c-kit and the HCN2 ion channel was co-expressed with c-kit.In the HCN blocking group,the green fluorescent labeled HCN1 and HCN2 ion channels were significantly reduced,and the ckit in the field of vision was also significantly reduced,despite the same treatment with moxibustion.It is suggested that moxibustion may regulate GI motility by regulating the number of active c-kit through HCN1 and HCN2 ion channels.HCN1 and HCN2 ion channels exist in the gastric antrum of the greater curvature,the upper small intestine,the satiation center and the lateral hunger center of the medial hypothalamus nucleus,and the increase and decrease of HCN1 and HCN2 ion channels in each region were synchronous in the experiment.It is suggested that HCN1 and HCN2 ion channel may be the material basis of synergism between brain and GI regulation of GI motility,and moxibustion can achieve the regulation of GI motility may be related to this way.【Conclusion】Moxibustion for the treatment of FGIDs is the best method to select various moxibustion methods in the experiment.Moxibustion can effectively regulate GI motility of FGIDs rats,which may be related to the up-regulation of HCN1 and HCN2 expression in gastric antrum,small intestine,satiety center and feeding center.