Correlation Between Tonggong Point And Mast Cells In Rats With Gastric Ulcer Model And Post-Translational Modification Of Gastric Tissue Protein After Electro-Acupuncture Intervention

Objective: With gastric ulcer model rats as the object of observation,under the condition of disease,used the immunofluorescence technique to observe the morphological and functional changes of mast cells in the "Tonggong" acupoint area and the changes of chemical components such as 5-HT,HA,CGRP,SP,etc,to explore the material basis of sensitization reaction of "Tonggong" acupoint and the effect of acupuncture;and to further observe the electro-acupuncture at Zusanli and Hemu acupoints.The difference in post-translational modification of gastric tissue protein between acupoints and stress gastric ulcer model rats can provide a scientific basis for acupoint selection and compatibility in future acupuncture treatment of stress gastric ulcer.Methods: Choose the "Tonggong " acupoint of epigastric pain,which has the highest frequency and is located at four acupoints of abdomen,back,upper limb and lower limb,namely Zhongwan,Weishu,Neiguan and Zusanli acupoint.Four other non-epigastric pain "Tonggong points" distributed in abdomen,back,upper limbs and lower limbs were selected as control points,namely Guanyuan,Ganshu,Quchi and Sanyinjiao acupoint.According to the experimental steps: anesthesia?select point?hemoperfusion?skin tissue removal?frozen section?immunofluorescence staining?sealing,to observe the positive expression of mast cells(MC)and calcitonin gene-related peptide(CGRP),serotonin(5-HT),histamine(HA)and substance P(SP)in skin tissue of 8 acupoints in normal rats and gastric ulcer model rats,and make statistical analysis.Furthermore,selected the commonly used to treat epigastric pain Zusanli acupoint,to stimulate the gastric ulcer model rats Zusanli acupoint by electro-acupuncture,and compared with model group to explore the relationship between the effect of acupuncture and acupoint sensitization.Finally,through the posttranslational modification technology,according to the experimental steps: protein extraction ?protein quantification ?electrophoresis ?transmembrane ?membraneclosure incubation of antibodies ?incubation of antibodies? exposure ?analysis of results,explore the difference of the effect of acupuncture at single acupoint and compatible acupoints on viscera at the protein level.Results: 1.In physiological state,MC content in local skin tissues of abdominal and dorsal acupoints is less,while that in local skin tissues of limbs is relatively higher;5-HT and HA exist widely in skin tissues,and often share positive expression with MC;CGRP positive expression is found in local skin tissues of acupoints of different parts: There are fewer in abdomen,fewer in back and more in limbs.There are differences in SP positive expression in local skin tissues of acupoints in different parts: abdomen and back are very few,limbs are relatively more;and acupoints with similar parts have the same chemical composition and quantity in local skin tissues.2.Compared with normal rats,the positive expression of MC,5-HT,HA,CGRP and SP at Zhongwan,Guanyuan,Weishu,Neiguan,Quchi,Zusanli and Sanyinjiao acupoints increased significantly,while the positive expression of MC at Ganshu acupoints did not change significantly,but the positive expression of 5-HT and HA at Ganshu acupoints increased significantly.Compared with the similar non-Tonggong acupoints,Zhongwan,Weishu,Neiguan and Zusanli have more positive expressions of MC,5-HT and HA in Zhongwan,Weishu and Zusanli,and the positive expression fibers of CGRP and SP in Zhongwan and Zusanli are longer.In addition,degranulation of MC was observed in the skin of rats with gastric ulcer.3.Compared with the positive expression of MC in Zusanli acupoint of gastric ulcer model rats,electro-acupuncture intervention on Zusanli acupoint of gastric ulcer model rats could enrich MC further,increase the number of MC,and increase the degranulation rate of MC;increase the positive expression of 5-HT and HA;increase the length of CGRP and SP fibers in skin tissue of the acupoint area.It shows that MC,5-HT,HA,CGRP and SP are the material basis for acupuncture to play its role.4.In the course of intervention of gastric ulcer model rats by electro-acupuncture at single acupoint and acupoint combination,significant differences can be seen in WB modified screening spectra between blank group,model group,single acupoint group and acupoint combination group,and they are significantly regulated by 2-hydroxy isobutyrylation,malonylation,succinylation,acetylation and phosphorylation.Conclusion: 1.Under the condition of disease,the "Tonggong Point" related to disease will have specific sensitization reaction.2.MC,5-HT,HA,CGRP and SP are the important material basis for acupoint sensitization.3.MC,5-HT,HA,CGRP and SP are important carriers for the effect of acupuncture and moxibustion.4.Acupuncture at single acupoint and compatible acupoints has different regulatory effects on viscera.