| Objective: To Study the gastric mucosal proteins expression in chronic gastictiswith damp-heat syndrome of spleen-stomach and investigate the pathogenesis relatedto the chronic gastictis.To observe the differential expression of gastric mucosalprotein in chronic gastictis with damp-heat syndrome of spleen-stomach aftertreatment with sanren decoction and investigate the mechanism of sanren decoction inchronic gastictis with damp-heat syndrome of spleen-stomach.Method:1.Study on establishment of chronic gastictis rat model with damp-heatsyndrome of spleen-stomach and treatment with sanren decoction:36male SpragueDawley rats were divided into three groups randomly,normal control group, modelgroup, sanren decoction group.The rat models were feeded with sweet and greasyfood,treated with damp heat environment,2%sodium salicylate and liquor weregavaged.The content of gastrins(GAS) and motilin(MTL) was detected byradioimmunoassay.The degree of inflammation was observed in gastric mucosal dyedby hematoxylin-eosin(HE) in each group.2.Study on proteomic analysis of gastric mucosal in chronic gastictis rat modelwith damp-heat syndrome of spleen-stomach and treatment with sanren decoction：gastric mucosal total protein was extracted from normal control group, model group,sanren decoction group respectively.Two-dimensional gel electrophoresis (2-DE)technique was used to separate gastric mucosal total protein.After coomassiestaining,the two-dimensional gel electrophoresis (2-DE) profiles were analysised todetect protein spots of differential expression by PDquest8.0software, the proteinspots of differential expression was acquired peptide mass fingerprinting(PMF) byMALDI-TOF/TOF-MS.Eight peaks was selected to process MS/MS.Both the MSand MS/MS data were integrated and processed by using the GPS3.6software andMascot2.1software,then the protein spots of differential expression was identified.Result:1.The chronic gastictis rat models with damp-heat syndrome of spleen-stomach were established by quantified methods successfully.The gastric mucosal inflammation was observed obviously through histopathology in model group, gastricmucosal inflammation was reduced after treatment with sanren decoction.The contentof GAS and MTL from model group was increased,compared with normal controlgroup and sanren decoction group,and the difference was statistically significant (p<0.05).2.The gastric mucosal two-dimensional gel electrophoresis profiles of normalcontrol group,model group and sanren decoction group were established with highresolution and good repeatability under the same conditions。The protein spots were1025±39,994±51,1087±33,deteced from two-dimensional gel electrophoresisprofiles of normal control group, model group and sanren decoction grouprespectively. The protein spots of differential expression were74between modelgroup and normal control group,30spots up-regulated in model group while44spotsdown-regulated; The protein spots of differential expression were75between sanrendecoction group and model group,49spots up-regulated in sanren decoction groupwhile26spots down-regulated.Five protein spots of differential expression of themost obvious were selected and identified by MALDI-TOF/TOF-MS after comparingwith the two-dimensional gel electrophoresis profiles of normal control group,modelgroup and sanren decoction group.The identificated results are: heat shock protein72,heat shock protein60,protein disulfide-isomerase,malate dehydrogenase,unnamed protein. Their function is mainly related to molecular chaperones andenergy metabolism.Conclusion: The method of feeding sweet,greasy and liquor,2%sodiumsalicylate under damp-heat environment was ideal to establish the chronic gastritismodel with damp-heat syndrome of spleen-stomach.Sanren decoction cansignificantly improve gastric mucosal inflammation,clinical symptoms and signs ofchronic gastictis with damp-heat syndrome of spleen-stomach,also can reduce thelevel of GAS and MTL.The gastric mucosal2-DE profiles of normal control group,model group,sanren decoction group were established successfully,and five proteinspots of differential expression were identified.The pathogenesis of chronic gastictiswith damp-heat syndrome of spleen-stomach may be related to energy metabolism disorders and stress,the mechanism of sanren decoction in chronic gastritis withdamp-heat syndrome of spleen-stomach may adjust the differential expression ofgastric mucosal protein. |
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