Study On The Effect Of Electro Acupuncture (EA) At Foot-Yangming Meridians On Related Protein Phosphorylation In The Recovery Of Gastric Mucosa With Protein-chip Technology

Objective:To study the effect of the stimulation of ST with EA on the injury repairing of gastric mucosa. By observing the changes of related signal protein phosphorylation under the stimulation of ST with EA, the related signal molecule and pathway were revealed, and from the proteome analysis, the mechanism of injury repairing of Stomach mucosa under the stimulation of ST with EA was expounded which applied certain experimental and theoretic basis for clinical applying and was helpful to understand the therapeutic mechanism of EA and the relationship between ST and stomach.Methods:40 SD rats were randomly divided into 4 groups:contrast group, model group, EA treating group and EA contrast group with 10 rats per group. The research was divided into three parts:Part 1:The stomach mucosa injury model was set by gastric administration with ethanol. The injury indexes of stomach mucosa were measured by GUTH. The morphology changes of stomach mucosa in 5 randomly-selected rats were observed by HE. The proliferating cell nuclear antigen (PCNA) was tested by immunohistochemical method. Part 2:The rest SD rats were randomly divided into 4 groups with 5 rats per group. The stomach mucosa was extracted. The changes of related signal protein phosphorylation of stomach mucosa injury repairing with EA at ST were detected by 720 kinds of phosphorylation of antibody protein chip tech. And the differential expressed phosphorylation signal protein in each group was screened. Part 3:the express of raf-1 protein of stomach mucosa in the samples of part 1 and 2 were detected with Western-blot and the expression of ERK 1/2 protein and cyclin D1 protein were detected by SP.Results:1. The number of the injury index of stomach mucosa in EA treating group was decreased obviously, and compared with model group and EA contrast group, the injury index was remarkable lower (P<0.01); compared with contrast group, the difference was insignificant (P>0.05); compared with contrast group, the index in model group was increased obviously and the deference was remarkable significant (P＜0.01).2. EA at ST can promote the proliferation of stomach mucosa cell. Compared with model group, the difference of the level of PCNA-LI in EA treatment group and model group was remarkable significant (P<0.01).3.720 kinds of phosphorylation of antibody protein chip tech detection shows: compared with contrast group,84 kinds of protein that was up regulated more than 1.5 folds in the phosphorylation change were up-regulated,12 kinds were down-regulated; while in EA contrast group, only 17 kinds were up-regulated and 3 were down-regulated; compared with EA contrast group, the number, function type and signal transduction pathway of phosphorylation signal protein were much better than that of EA contrast group.4. EA at ST can obviously activate the level of RAF-1 and phosphorylation of protein and up regulated the level of cyclin D1 protein. The difference among the groups were all remarkable significant (P＜0.01 or P＜0.05).Conclusion:1. EA at ST can obviously improve the morphology changes of stomach mucosa and increase the index of PCNA-LI which indicates that EA at ST can obviously improve the morphology changes of stomach mucosa2. EA at ST can dual-directionally regulate the proteins of model group which involves various functional types and belongs to multiple signal transduction pathways. It indicates multi-target and multiple-way is the important characteristic of EA at ST to repair the stomach mucosa.3. Compared with contrast group, EA at ST can activate the MAPKS signal transduction pathway specifically which indicates MAPKS signal transduction pathway is close related with repairing of EA at ST to promote the repairing of stomach mucosa.4. EA at ST can obviously activate various functional types of proteins and several signal transduction pathway which accord with the basic characteristics of acupuncture such as holism and comprehensibility. Studying the holistic regulating effect of TCM and acupuncture by using protein chip tech can promote the understanding and applying of holism in TCM.