A Study Of Electroacupuncture On EGC Cell Morphology And GDNF Expression In Functional Constipation Mice

Objective:As the research intended to use the functional constipation animal models as the carrier,To explore the mechanism of electroacupuncture Back-shu and Front-mu points for treating functional constipation by EGC and GDNF, Which are the keys in the process of gastrointestinal.Methods:Choose 64 KM mice of clean level, half male and half female, randomly divided them into blank control group, model group, acupuncture group and EGC group, each group had 16 mice,1 week is a period of treatment, each group of animals divided into two subgroups according to the treatment period, each of the subgroup has 8 mice, to take samples after accepted acupuncture therapy for 1 week and 2 weeks. Control group recieved 0.9% saline with the dose of 10 mg/kg·d to intragastric administration, the capacity is 0.1 ml/10 g, lasted for 14 d. Model group、acupuncture group and EGC group recieved the mixed suspension of Compound Diphenoxylate Tablets with the dose of 10 mg/kg· d to intragastric administration, the capacity is 0.1 ml/10 g, lasted for 14 d. After the FC mould finshed, control group and model group just fixed without acupuncture, acupuncture group and EGC group aceepted acupuncture therapy for 1 week and 2 weeks, EGC group should inject EGC inhibitors in abdomen before acupuncture therapy.Choose the colon tissue after treatment, observed the morphological changes of colon tissue and EGC cell by using optical and electron microscope,and TUNEL staining to detect apoptosis count, the in situhybridization combined with technique methods of computer image analysis and immunohistochemical, detected GDNF mRNAand protein expression, comparing differences among groups.Results:1. Histomorphology result:There were pathomorphological changes in different degree of colon tissue in the model group,acupuncture group, the inhibition of EGC group,such as edema of nerve in mucosal,degeneration of EGC cells,infiltration of inflammatory cells,pathological changes was least in the acupuncture group;2. TUNEL apoptosis detection results:Through statistical analysis on the positive rate of TUNEL apoptotic cells, the difference was statistically significant in acupuncture group and model group (P<0.05);from the observation of the morphology and cell apoptosis,there was no significant difference in mice which received different periods of acupuncture; morphology has improved in animal model with the inhibition of EGC cells after acupuncture,but there are still significant differences compared with the acupuncture group (P<0.05).3. GDNF protein expression results:The expression of GDNF protein and mRNA in model group was significantly decreased (compared with control group,P<0.01).EA back-Shu and front-Mu points can improve the expression of GDNF protein and mRNA (compared with the model group, P<0.05),light microscope and electron microscope found that acupuncture can repair damage depithelial cells, and then improve the intestinal transmission function. EGC group in the application of EGC inhibitor destructing EGC cells before EA stimulation,showed the expression of GDNF protein inthe colon had no significant difference compared with the model group (P>0.05), compared with the EA group significantly decreased (P<0.05).4. GDNFmRNA expression results:The expression of GDNFmRNA was higher than that in the model group (P<0.05), but its level of expression was lower compared with the acupuncture group, but the difference is not significant (P>0.05). The morphology compared with acupuncture group was more serious.Conclusions:1. EA back-Shu and front-Mu points can improve the expression of GDNF protein and mRNA secreted by EGC cells,repair the colonic epithelial cells, and then improve the intestinal transmission function.2.Afeter the EGC cell had be destoried,the expression of GDNF had decereased.The incereasing of EA to GDNF was no longer significant,the repairation after colon injury becamed slowly.3. Different period of therapy to colon tissue and EGC cells of animal model made no significant difference. And their effcetion to expression of GDNF and mRNA had no significant difference.either.