| ObjectiveUnder physiological condition,using a new generation of highly sensitive retrograde tracer to observe the regulation and relevance of the peripheral and central nervous system on the Yuan points,Luo points,He points,Shu points,Mu points of the stomach meridian of Foot Yangming.And elucidate the internal relationship between the stomach and its related specific points from three aspects of sensation,movement and autonomic nervous system.To lay a neuroanatomical foundation for the mechanism of specific acupoints in the treatment of visceral diseases.Taking the stress gastric ulcer model rats as the research object,by using the method of immunofluorescence chemical staining,observing the morphological and functional changes of vasoactive intestinal peptide(VIP)and substance P(SP)in gastric tissue of rats with gastric ulcer model after acupuncture combination of He and Mu points,combination of Yuan and Luo points,combination of Shu and Mu points.From the molecular level to explain the pathological mechanism of the treatment of gastric ulcer.It lays a material foundation for acupuncture to intervene the mechanism of viscera in physiological and pathological state.It provides a new model or direction for the study of the correlation between acupoints and viscera.It guides significance for acupuncture and moxibustion to select a reasonable compatibility scheme for the treatment of corresponding visceral diseases.MethodsPart I:Thirty six clean male Sprague Dawley rats with a body mass of(220±20)g were randomly divided into these groups: He point BL21 and Mu point CV12,Shu point ST36 and stomach,Yuan point ST42 and Luo point ST40,.When the rats were under respiratory anesthesia,0.1% of AF488-CTB or AF594-CTB were injected into "CV12","BL21" and "ST36","ST40" and "ST42" points on the left side of the corresponding six groups.After tracing 2-3 days,anesthesia and cardiac perfusion.Removing The bilateral sympathetic nerve chain,DRG,spinal cord,brain stem and upper and lower segments of vagus nerve.The removed tissue was placed in the fixed solution containing 4% paraformaldehyde and fixed for 2-3 hours.After cleaning with 0.1 mol/L PB,it was dehydrated in 0.1 mol/L PB containing 25% sucrose and placed in the refrigerator at 4℃,and then the tissue sank naturally.The labeled neurons were observed and recorded by fluorescence microscope or confocal microscope.Part II:Thirty clean male Sprague Dawley rats with a body mass of(220±20)g were randomly divided into five groups: Blank group A,Model group B,combination of He and Mu points group C,combination of Yuan and Luo points group D,combination of Shu and Mu points group E,six in each group.The rats were under respiratory anesthesia and electroacupuncture.After 7 days,the pathological model of gastric ulcer was made by water immersion binding method.Anesthesia and cardiac perfusion.Take out the stomach,fix it for 2-3 hours,then change it to 0.1 mol/L PB(pH7.4)containing 25% sucrose for dehydration,and then place it in the refrigerator at 4℃ for the tissue to sink naturally.The sections were made into 20μm thick sections by frozen microtome,which were pasted on the positiveion glass slide,stained vasoactive intestinal peptide and substance P with immunofluorescence staining technology,and then observed and recorded with fluorescence microscope.ResultsPart I:1.The sensory neurons labeled by AF594/488-CTB related to "Stomach" and "BL21" on the left appeared in the dorsal root ganglion of T2-14 and T8-12,and concentrated in T8-11 section.The motoneurons labeled by AF488-CTB innervating "BL21" area were distributed in the left anterior horn of T11-13 segment.The sympathetic postganglionic neurons,which dominate the "Stomach" and "BL21",are mainly distributed in the left thoracic sympathetic nerve chain.Labeled neurons were observed in the dorsal nucleus and the superior and inferior segments of the vagus nerve.2.The sensory neurons labeled by AF594/488-CTB related to "ST36" and "CV12" appeared in the dorsal root ganglion of L3-L6,T6-T13 segments respectively.The motoneurons innervating the "ST36" and "CV12" were distributed in the left anterior horn of L3-L5 andT8-T12 section respectively.The sympathetic postganglionic neurons in "ST36" and "CV12" acupoints are mainly distributed in the left lumbar sympathetic chain and the lower thoracic sympathetic chain.3.The sensory neurons labeled by AF594/488-CTB related to "ST40" and "ST42" appeared in the dorsal root ganglion of L3-L6 segments respectively.The motoneurons of "ST40" and "ST42" were distributed in the left anterior horn of L4-L6 respectively.Part II:1.In the model group,the gastric mucosa was thickened,the inner wall was red,and there were many large black and brown bleeding spots.In the combination of He and Mu points intervention group,the gastric mucosa was slightly thickened and the inner wall was slightly red with occasional bleeding points.In the combination of Yuan and Luo points intervention group,the gastric mucosa was slightly thickened,the inner wall was pink and there were many bleeding spots.In the combination of Shu and Mu points intervention group,the gastric mucosa was slightly thickened and the inner wall was slightly red with occasional bleeding spots.2.The length of VIP positive nerve fibers in the gastric wall of the Model group was significantly lower than that of the Blank group(P(27)0.01).After acupuncture intervention,the length of VIP positive nerve fibers in gastric wall also increased in varying degrees:Among them,there was a significant increase in the length of the combination of He and Mu points intervention group compared with the Model group(P(27)0.01),and close to the blank group(P(27)0.01);There was a significant increase in the combination of Shu and Mu points intervention group compared with the Model group(P(27)0.01),and close to the Blank group(P(27)0.01);Compared with the Model group,the combination of Yuan and Luo points intervention group increased,but there was no statistical difference(P(29)0.05);There was significant difference between the combination of Yuan and Luo points intervention group and the Blank group(P(27)0.01);There was no significant difference between the He and Mu points intervention group and Shu and Mu points intervention group(P(29)0.05).There was a significant increase between the Yuan and Luo points intervention group and the He and Mu points intervention group(P(27)0.05).There was an increase in the intervention group of the Yuan and Luo points intervention group and the Shu and Mu points intervention group,but there was no significant difference(P(29)0.05).3.The length of SP positive nerve fibers in gastric wall of Model group was significantly longer than the Blank group(P(27)0.01).After acupuncture intervention,the length of SP positive nerve fibers in the gastric wall also decreased in varying degrees: the intervention group of He and Mu points was significantly lower than the Model group(P(27)0.01),and close to the Blank group(P(29)0.05);the intervention group of Shu and Mu intervention group was significantly lower than the Model group(P(27)0.01),and close to the Blank group(P(29)0.05);the intervention group of Yuan and Luo intervention group was significantly lower than the Model group(P(27)0.05),but there was significant difference than the Blank group(P(27)0.01);There was no significant difference between the intervention group of He and Mu point with Shu and Mu intervention group(P(29)0.05);the intervention group of He and Mu points was significantly lower than the Yuan and Luo intervention group(P(27)0.05);the intervention group of Shu and Mu points was significantly lower than the Yuan and Luo intervention group(P(27)0.05)Conclusion1.In the sensory innervation of the stomach and its specific points,there are different ganglion segments in the stomach and its specific points.2.BL21,ST36 and CV12 can regulate the stomach in different degrees through the conduction of sympathetic nerve.3.Acupuncture at specific points of the stomach will not affect the stomach through the motor conduction pathway.4.In physiological state,Shu and Mu points are compatible with the proximal ganglia of the stomach,which can play a synergistic role in sensory,motor and sympathetic pathways.The Yuan and luo points are compatible with the distal ganglia of the stomach,play a synergistic role in the control of sensory and motor neurons.He and Mu is the compatibility of the distal and proximal segments of the stomach,it may play a regulatory role in the stomach through the stimulation of the advanced center..5.Under pathological condition,the combination of He mu,Shu Mu and Yuan Luo can prevent and treat the stomach by adjusting the secretion of VIP / SP in gastric tissue.And the effect of the combination of He and Mu is the best. |
