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1. Establishment of experimental chronic atrophic gastritis rats model by MNNG and other factorsObjective At present commonly used MNNG to establish chronic atrophic gastritis model in rats exists generally time consuming, big cost, formed model dose not unified, different time of appearing pathological changes and other issues, and all most lack of observation whether it can recover after stoping making model. This experiment try to establish experimental chronic atrophic gastritis (CAG) model in rats by given MNNG, ranitidine, alcohol and hunger satiety disorders. Explore the process dynamically by MNNG and other factors.Dynamic observing the pathological process of chronic atrophic gastritis and the self recovery situation after stoping making model4weeks and8weeks.Methods①Observing high and low two different concentrations MNNG solution loading multiple factors to build CAG rats model:Male Wistar rats were randomLy divided into3groups, including the control group, model1group (80μg/mLMNNG group) and model2group (150μg/mLMNNG group). Rats in the control group were fed with normal food, while rats in the model1and model2groups were given MNNG solution (modell:80μg/mLMNNG, model2:150ug/mL MNNG) every day;ig ranitidine0.03g/kg rats body weight; rats fasting14-16hours1times a week, ig45%alcohol after fasting,not given ranitidine that day.Continuous modeling for16weeks.②Observe the CAG rats gastric mucosa lesions recovery situation after stoping making model:16weeks later,model1group and model2group rats were randomLy divided into2groups each, including model A group (Recovery group) and model B group(keep modeling group). In the20week killed part of each group rats, removed the gastric mucosa tissue and soaked in10%formalin liquid, HE staining, analysis the pathological results. In the24week, killed all rats, removed the gastric mucosa tissue and soaked in10%formalin liquid, HE staining, observed the gastric mucosa lesions.〥ynamic monitoring of gastric mucosa lesions in different period of CAG in rats:weekly observation each group rats eating, drinking and activity status; observe the rats gastric acid secretion and gastric mucosa lesions by HE staining in12,16,20and24week.Results①Model1and model2group rats eating and drinking during modeling were significantly decreased, weight growth was slower.②In12week model1and model2group rats had appeared atrophic gastritis pathological changes, model2rats got more serious lesions than modell rats.③In16week model1and model2group rats were markedly atrophic gastritis pathological changes, parts associated with intestinal metaplasia in the model2groups. Model B group rats gastric mucosa lesions continued to worsen,accompanied by intestinal metaplasia in20-.24week.④Recovery4weeks and8weeks,M1-A and M2-A group rats gastric mucosal lesions did not see obvious self-healing phenomenon,M2-B group rats associated with intestinal metaplasia and hyperlasis. Recovery8weeks M1-A group rats body weight growth obviously, close to the control group.⑤In12,16,24week, the rats gastric acid results can’t reflect the real situation due to the measurement method not mastered.Specific see experiment1discussion section.Conclusion①In summary, experimental chronic atrophic gastritis rats model can be established by MNNG with other factors.②Rats drink80μg/mL MNNG and150μg/mL MNNG solution can form chronic atrophic gastritis at the12week,drink150μg/mL MNNG solution with other factors can emerge intestinal metaplasia at the16week,and gastic precancerous lesions at the20-24week.③The CAG rats model gastric mucosa pathological changes did not see obvious recovery when stop making model for4weeks and8weeks.Only80μg/mL MNNG CAG model group rats weight growth had obvious recovery but not150μg/mL MNNG CAG model group.2. The experimental research of SQ granule on the treatment of chronic atrophic gastritisObjective SQ granules were professor Jianhua-Dong’s experience prescription, was hospital preparations and clinical used in the treatment of chronic atrophic gastritis.It made up of9taste traditional Chinese medicine.Now it was made into granules throμ gh modern medicine technology.The purpose of this part is to establish experimental chronic atrophic gastritis(CAG) model in rats refer to the first part and explore the effect of SQ granules on rats general condition,gastric mucosal lesions, provide the basis experimental data for its clinical curative and new drug application.Methods CAG model was established by120ug/mL MNNG solution MNNG and other factors for14weeks with reference to the first part.Then the model rats were divided into6groups randomLy, which were ig administered with saline, Moluodan,folic acid,SQ granules18、9and4.5mg/kg,respectively,taking normal rats as control group at the beginning of modeling.Monitoring each group rats general conditions.In22week, gastric tissue for routine HE staining, calculation of mucous membrane thickness/muscle layer thickness (L1/L2).Results①The CAG rats’weight were obviously lighter than the control group, the SQ granules and folic acid group could increase the body weight of CAG rats.②SQ granules in each dose group could obviously improve the elimination of chronic atrophic gastritis rats gastric mucosa pathological changes, stomach wall thicker, flexibility was better, had a reverse effect in intestinal metaplasia and dysplasia hyperplasia,decreased the thickness of the mucosal muscularis,increased the mucous membrane of the gastric mucosa thickness/muscle layer thickness ratio.③SQ granules in each dose group could improve the gastric acid secretion and Pepsin activity.Conclusion SQ granules had a significantly effect on improving gastric mucosa lesions of CAG model rats. It decreased the thickness of the mucosal muscularis, increased the mucous membrane of the gastric mucosa thickness/muscle layer thickness ratio, improve the gastric acid secretion and Pepsin activity. Indicated that SQ granules had obvious intervention effect for chronic atrophic gastritis, showed a good therapeutic effects.3. The mechanism research of SQ granules on CAG ratsObjective Etiology and pathogenesis of chroic atrophic gastritis were not clear yet, and there were still no very effective treatment methods and specific western medicine. Traditional Chinese medicine had made gratifying progress for clinical treatment of CAG, but different treatment methods, treatment mechanism is also different.In this part,review the latest literatures, try to discuss the mechanism of SQ granules cureing chronic atrophic gastritis through gastrointestinal hormone, cytokines and COX-2pathway.Methods Grouping of experimental animals and dosing see the second part.〥etected the gastrointestinal hormone PGE2, SS, CCK, MTL, GAS, VIP and cytokines IL-1, GH, EGF by radioimmunoassay; GAS、SS、CCK have have the function of nutrion, protect the gastric mucosa; MTL、VIP、CCK、SS have the function of adjusting gastrointestinal tract secretion; GAS、SS、CCK have the function of adjusting gastric bowel movement;cytokines EGFand GH can promote gastric mucosa cell update repair, protect the gastric mucosa;IL-1is not only the important proinflammatory factor,and it is the strongest gastric acid secretion inhibitor.②Detected PGI and PG II by enzyme immunoassay; PGI/PG II reflect the gastric mucosal barrier function.③Detected COX-2、Bcl-2、IL-10and PCNA by immunohistochemistry. COX-2regulates the prcess of arachidonic acid metabolism and plays an important role in the development of CAG and GC. COX-2regulates the relevant signal factors on its path such as Bcl-2, IL-10and PCNA. Bcl-2inhibitor the cell apoptosis, PCNA reflects the cells proliferation status, IL-10is an immune inhibitory factor.Results①SQ granules in each dose group could improve the level of serum EGF, promote gastric mucosa epithelial cell growth, increase the gastric mucosal blood flow, protect gastric mucosa;②Improve the level of serum GH, promote the mucosal repair;③Improve the level of serum cck, regulate the movement of the gastrointestinal tract, and promote the secretion of various digestive enzymes;④Reduce the levels of serum GAS.⑤Model group rats cox-2,bcl-2were higher expressed,and SQ granules each dose group could significantly inhibit cox-2,bcl-2expression.Conclusion SQ granules have obvious intervention role of gastrointestinal hormone and cox-2pathway, inhibit atrophic gastritis to develop in the direction of precancerous lesions and even canceration.
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