| Objective:To observe the dynamic effect of moxibustion on metabolic profiles in biological samples(stomach,cerebral cortex and medulla)in Acute Gastric Mucosa Lesions in Rat,discussion the mechanism of Moxibustion treatment on Acute Gastric Mucosa Lesions in metabolic level.Method:all rats were randomly separated into four groups:a)control group(group A),b)gastric mucosal lesions(GML)model group(group B),c)GML rats with moxibustion treatment on the stomach meridian of Foot-Yangming(SMFY)acupoints(group C),and d)GML rats with moxibustion on non-acupoints(group D).All rats except for controls were randomly divided into three subgroups(n=6,per subgroup).In the first subgroup,rats were executed at 1d after treatment.As mentioned above,the rats from second and third subgroup was sacrificed at 4d and 7d,respectively.All rats except for controls were established GML modeling with alcohol given by gavage with 75%ethanol solution at 4ml/kg after deprived of food for 24h.In this study,we applied ~1H NMR-based metabolomics to dynamic analysis of metabolic profiles in biological samples(stomach,cerebral cortex and medulla).And the conventional histopathological examinations,the lesions index as well as metabolic pathways assays were also performed.Results:1.General Behavioral Observation of Rats:Compared with the normal group,the rats in the model group recovered to a certain extent in the later stage of the experiment because of their mental malaise,poor flexibility and reduced activity.Compared with the model group,the rats in the moxibustion group recovered faster,and the state of the moxibustion stomach meridian group recovered better than the control group.2.Histological observation of gastric mucosa:Compared with the normal group,the model group,the moxibustion stomach meridian group and the moxibustion control group had obvious congestion points on the surface of gastric mucosa at 1 day and 4 days,the wrinkles of gastric wall disappeared,the index of gastric mucosal injury increased significantly(all P<0.01 or P<0.05),and the histomorphology of gastric mucosa had significant pathological changes under light microscopy;at 7 days,the surface of gastric mucosa in the third group had no obvious congestion points by naked eye observation.Microscopic Mucosal pathological changes were improved to varying degrees.Compared with the model group,the structure of gastric mucosa in the subgroup of moxibustion at 4 days of stomach meridian and the control point recovered more completely than that in the model group,and the index of mucosal injury in the rats decreased(P<0.01,P<0.05).The subgroup of moxibustion at 4days of stomach meridian was also lower than that in the subgroup of moxibustion at4 days of control point(P<0.05).The injury of gastric mucosa in the moxibustion on stomach meridian group improved most3.Comparison of brain and stomach metabolic profiles of rats:(1)Gastric tissue:In 1 day,4 days and 7 days,GML model group was significantly separated from normal group,moxibustion group of stomach meridians and moxibustion control point group.The interval separation degree between moxibustion at stomach meridian points group and moxibustion control point group was poor.(2)Cerebral cortex:In 1,4 and 7 days,GML model group was separated from normal group,moxibustion at stomach meridian points group and moxibustion control point group in varying degrees,and the separation degree was better in the first and fourth days.Moxibustion at stomach meridian points and moxibustion control points can be distinguished at three time points.(3)Medulla oblongata:In 1 day,4 days and 7 days,GML model group was separated from normal group,moxibustion at stomach meridian points group and moxibustion control point group in varying degrees,and the degree of separation was better on the first day.The difference between moxibustion at stomach meridian points group and moxibustion control point group was most obvious on the fourth day.4.Metabolic markers in gastric tissue rats treated by moxibustion:(1)15metabolites in gastric tissues,including:beta-glucose,alpha-glucose,acetic acid,citric acid,pyruvic acid,oxaloacetic acid,isoleucine,leucine,valine,lysine,glutamic acid,glutamine,aspartate,tyrosine and phenylalanine.(2)8 metabolites in cerebral cortex,including succinic acid,alpha-ketoglutarate,creatine,glycerol phosphocholine,fumaric acid,taurine,glutamic acid and glutamine.(3)7 metabolites in medulla tissue,including lactic acid,citric acid,methionine,glycine,serine,glutathione and phenylalanine.5.The effects of moxibustion on the metabolic pathways of various tissues and the whole brain-stomach metabolic pathways in GML rats:(1)Moxibustion regulates13 metabolic pathways in gastric tissue of GML rats,mainly involved in energy metabolism,amino acid and amide metabolism.(2)Moxibustion regulates metabolic markers in the cerebral cortex of GML rats,involving eight pathways,mainly in the metabolism of amino acids and amides.(3)Moxibustion regulates eight pathways of metabolic markers in the medulla oblongata of GML rats,mainly involved in the metabolism of amino acids and amides.(4)Moxibustion regulates the whole brain-stomach metabolism of GML rats,involving 20 metabolic pathways,mainly involved in the metabolism of amino acids and amides.Conclusion:1.The results of naked eye observation,pathological observation and gastric mucosal injury index showed that the gastric mucosal tissue of rats had certain self-repairing ability to acute mucosal injury.Moxibustion could promote the repair of gastric mucosal injury and improve the morphology of the injured tissue.Moreover,moxibustion of stomach meridian points has a faster onset time and better healing of mucosal injury than moxibustion control points.2.Based on ~1H-NMR metabolomics technique,it reveals that the dynamic changes of metabolites in brain and stomach tissues during acute gastric mucosal injury in rats.The changes of metabolic markers at different time points indicated that rats,during the period of injury and repair about acute gastric mucosal,could be observed the disturbance in the energy metabolism,antioxidant capacity,immunity,cellular activity and neurotransmitter function of the brain and stomach tissues,and appear some dynamic changes of self-regulation relative with self-healing.3.Appling ~1H-NMR metabonomics conbined with multivariate data analysis,it reveals that moxibustion treatment on GML rats through earlier and longer term to promote energy release,enhancecell activity in target organ,inprove antioxidant activity,immune function and energy metabolism in brain tissues,and regulate metabolites and pathways related to neurotransmitter function in brain-stomach integrity,then futher to has beneficial effects on acute gastric mucosal lesions.4.Both two kinds of moxibustion treatments have different degrees in regulating the metabolic markers.Moxibustion at stomach meridian acupoints is superior to moxibustion control points in regulating brain-stomach energy metabolism and cell activity in GML rats.Moxibustion at stomach meridian acupoints is faster and longer curative effection,and it showed a specificity in meridian acupoints. |
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