The Mechanism Of Gastritis Ⅰ Regulating To The Component Of Extracellular Matrix In Stomach Of The CAG Rats

Objective:Chronic atrophic gastritis(CAG) is a refractory disease of digestive system diseases,which's main pathological changes including gastric mucosa and gastric glands thinning,lamina muscularis mucosae thickening,intestinal metaplasia,atypical hyperplasia, etc.According to the linkage theory of gastric cancer carcinogenesis,the pathological changes of gastric cancer carcinogenesis follow the step from chronic superficial gastritis (CSG),chronic atrophic gastritis(CAG),intestinal metaplasia,atypical hyperplasia and the end to gastric cancer.CAG as a kind of lesion precancerous has been generally accepted. Modern medicine can cure gastritis by eradicating HP,anti gastric acid secretion and providing nutritional support.But as to the pathological changes of atrophic gastritis, intestinal metaplasia and atypical hyperplasia,it doesn't have active drug to cure.But Tradition Chinese Medicine(TCM) according to the entirety theory to zheng differentiation-treatment can receive excellent curative effect in curing CAG.It has very important Science significance and social benefit to lucubrate the mechanism of TCM treatment to CAG.My tutor professor Liu Youzhang according to the Clinical symptoms of CAG,treat it as stomach painful abdominal mass of TCM.The basic pathogenesis include weakness of the spleen and stomach,dys-splenism,stomach downward propelling disorders,insufficiency of vital energy and blood,middle energizer disorder of vital energy and ascent-descent disorder,.Therapeutically,professor Liu Youzhang treat CAG by strengthening spleen,benefiting vital energy,nourishing blood,regulating stomach and promoting granulation.GastritisⅠis an effective prescription which was found by Professor Liu Youzhang in his several decades clinical practice.It has the effectiveness of strengthening spleen,benefiting vital energy,nourishing blood,regulating stomach and promoting granulation,and has been confirmed it's effectiveness in reconditioning gastric mucosa and resisting epithelial metaplasia of CAG.The purpose of our empirical study is to lucubrate the mechanism of gastritisⅠcuring the CAG rats by lucubrating the mechanism of gastritisⅠregulating to MMP1,TIMP1 and integrin-β1 in stomach of the CAG rats,and to provide scientific experimental evidence of the clinical therapeutic effect of gastritisⅠ.Our empirical study for the first time lucubrate the interrelation of mRNA expressing and enzymic activity between MMP1 and TIMP1,and their synergy in the mechanism of the CAG rats.Our empirical study also researches the mechanism of cell signal transduction between extra cellular matrix(ECM) and cell adhesion molecule of the CAG rats.Methods:Our research was divided into overview and empirical study.It doesn't have the CAG disease name in TCM.So in the overview,the first we unscrambled the corresponding disease name in TCM to the CAG.We expounded the etiological factor, pathogenesis and therapeutic principle of ancient and contemporary doctor of TCM.We also expounded the advancement of modern medicine.In the empirical study,84 healthy SD rats were divided into 6 groups,including model group(24),normal group(12), western medicine control group(12),and 3 GastritisⅠtreatment group including high dose group(12),medium dose group(12),low-dose group(12).the CAG model rats was constructed by freely drinking the MNNG solution(50μg/ml),being intragastric administration and irregular diet.After the CAG model was constructed successfully, giving 5g/(kg·d) GastritisⅠto high dose treatment group,giving 2.5g/(kg·d) GastritisⅠto medium dose group,giving 1.25g/(kg.d) GastritisⅠto low-dose group,giving 0.86 g/ (kg.d) Wei Mei Su Pian to western medicine control group,the normal group was feed normally.We adopted gelatinase zymography to detect MMP1 enzymatic activity,inverted gelatinase zymography to detect TIMP1 enzymatic activity,fluorescent quantitation PCR to detect MMP1 and TIMP1 mRNA expression,adopted immunohistochemistry to detect integrinβ1 expression.Result:①Result of light microscope observe:The changes of gastric mucosa of CAG model rats included structure disorder,comprehensive glandular organ thinningz and atrophy and Saccular ectasia,the intestinal metaplasia and atypical hyperplasia dysplasia hadn't develop jet.The gastric mucosa of normal rats was normal.The gastric mucosa of the CAG rats treated by medium dose gastritisⅠwas line up in order,and glandular organ thinningz and atrophy and Saccular ectasia,the intestinal metaplasia and atypical hyperplasia dysplasia hadn't develop jet.The gastric mucosa of the CAG rats treated by low dose gastritisⅠwas structure disorder,major glandular organ thinningz and atrophy and Saccular ectasia.The gastric mucosa of the CAG rats treated by high dose gastritisⅠwas slightly structure disorder,a little glandular organ thinningz and atrophy and Saccular ectasia,and slightly necrosis.The gastric mucosa of the CAG rats treated by Wei Mei Su Pian was slightly structure disorder,a little glandular organ thinningz and atrophy and Saccular ectasia.②The result of fluorescent quantitation PCR found that the MMP1 mRNA expression of the CAG model group rats was lower than normal control group (P＜0.05).After the treatment of gastritisⅠ,the curative effect of medium dose gastritisⅠwas better than low dose and Wei Mei Su Pian(P＜0.05);the result of gelatinase zymography found that the MMP1 enzymatic activity of the CAG model group rats was higher than normal control group(P＜0.01),after the treatment of gastritisⅠ,the MMP1 enzymatic activity of medium dose and high dose gastritisⅠwere lower(P＜0.001),but the MMP1 enzymatic activity of western medicine control group and low-dose group were not obviously change(P＞0.01),the curative effect medium dose gastritisⅠwas best.③Result of fluorescent quantitation PCR found that the TIMP1 mRNA expression between the CAG model group rats and normal control group wasn't significant deviation(P＞0.05), and it was the same between the groups after the treatment(P＞0.05).Result of inverted gelatinase zymography found that the TIMP1 enzymatic activity of the CAG model group was lower than normal control group(P＜0.01).After the treatment of medium dose gastritisⅠ,the TIMP1 enzymatic activity was higher(P＜0.01),and didn't have significant deviation with normal dose(P＞0.05).Wei Mei Su Pian also had effect to raise the TIMP1 enzymatic activity(P＜0.05),but the curative effect medium dose gastritisⅠwas best.④Result of quantitative immunohistochemistry found that the integrinβ1 expression average coloration and average coloration density of the CAG model group was higher than normal control group(P＜0.01),after the treatment of medium dose gastritisⅠ,the integrinβ1 expression average coloration and average coloration density were lower obviously(P＜0.01),and didn't have significant deviation with normal dose(P＞0.05).The high dose and low-dose gastritisⅠalso had effect to raise the integrinβ1 expression average coloration and average coloration density,but the curative effect of medium dose gastritisⅠwas best.Conclusion:①Our study adopted synthetic methods to construct the CAG model rats. Those methods were able to build the CAG model rats successfully,but the reagents of this method was harmful to human body and environment,the time of intragastric administration was too long,that would be increase death rate.Therefore,we can explore better methods of modeling in the future experiment.②In our experiment,we simultaneously studied the interrelation of mRNA expression and enzymatic activity between MMP1 and TIMP1 for the first time.The result found that MMP1 and TIMP1 participates the development of the CAG model rats in the enzymatic activity level.The mechanism of action was adjusting the repair and regenerate of ECM by regulating MMP1 and TIMP1 enzymatic activity.③The result of our study found that integrinβ1 participates the development of the CAG model rats.The mechanism of action was that the long-term damage and reparative process of the CAG rats made the expression of integrinβ1 increase, the adhesions between cells was strengthened,and transfer various kinds of growth and cell differentiation information by signal transduction of cells,and the end to promote the regeneration of stomach mucous membrane.④The result of our study found that gastritisⅠhad satisfactory curative effect and renovation to the damage of the stomach of the CAG rats.It could promote inflammation recovery and deteriorate glandular organ atrophy of the CAG rats,the curative effect of medium dose gastritisⅠwas the best.