Mechanism Of Flavored Before The Zuojinwan Treatment Of Gastric Lesions In Rats

ObjectiveTo construct the model of Precancerous lesions of gastric cancer(PLGC) in rats by Simulating human etiological factor, observing the remedial effect of Jia Wei Zuo Jin Wan(JWZJW) on rats with PLGC and study the mechanism of JWZJW on PLGC at molecular level. Methods1. The way of constructing modelWe constructed model in a modified synthetic way as follow: we let the rats freely drink 100ug/ml MNNG, eat pure granula feedstuff including 0.03% Ranitidine, take a dose of 56℃ 15% NaCl by 10ml/kg by intragastric administration in the morning. We fed the rats for every two days and forbidded them from eating on the every third day . In the feeding two days, the rats took a dose of 20mmol (0.85%) sodium deoxycholate by 10ml/kg in the afternoon of feeding, on the fasting day the rats took a dose of 40% alcohol by 10ml/kg in the afternoon. The tails of rats were Clamped once a week for 30 minutes .The above-mentioned way was kept for 24 weeks. two rats were respectively chosen from the normal group and the model group at the end of the 24th week and then killed to judge if the model was successfully made according to histopathology . 2. Animal groupingSix-week-old male SD rats were used for the experiment. A total of 60 rats were randomly divided into the normal group(n=10) and the model group(n=50),in which 43 rats survived . These 43 rats were then randomly divided into model control group(n=11), tretinoin group (n=8), high-dose JWZJW group(n=8), middle-dose JWZJW group(n=8),low-dose JWZJW group(n=8).3. Administration dosageAfter the end of 24th week ,normal group was without any treatment ,Distilled Water was given intragastricsily to the rats in model group by 10ml/kg, various densities (200% , 100% , 50%) of JWZJW were given intragastricsily to the rats in high-dose , middle-dose , low-dose JWZJW group by 10ml/kg, tretinoin was given to the rats intragastricsily in tretinoin group by 10ml/kg. once a day for 16 weeks.2. Observation items(1)general conditions: to observe eating ,drinking, mentality, physique , activity, clothing hair, stool and mortality. (2)body weight: the rats were weighed once a month from the beginning of the experiment, and their mean weight was calculated and analyzed statistically.(3)gastric mucosa state under the macroscopic observation: to observe the color and luster .wrinkles and extensibility of the gastro-wall, inflammation before and after administration .(4)gastric mucosa state under light microscope observation:: to observe the change of gastric mucosa in histopathology (including lump, JM, ATP, inflammation, atrophy which were pathologically diagnosed in accordance with classifying and diagnostic standtard of experimental gastric adenocarcinoma ) and count their number. The dead rats in experiment wereexcluded in histopathological and statistical analysis. (5)To detect the expression rates of Proliferating Cell Nuclear Antigen (PCNA) by immunohistochemistry method and calculate Proliferating index(PI).(6)To detect the expression rates of oncogene protein in Epidermal Growth Factor Receptor (EGFR)> Vascular endothelial cell growth factor (VEGF), Hepatocyte growth factor receptor (C-met),(Bcl-2) by immunohistochemistry method.(7)To detect the expression rates of Cyclooxygenase 2 ( COX-2 ) and human telomerase reverse transcriptase (h-TERT ) by immunohistochemistry method.(8)Tb detect apoptosis cell by Terminal deoxynucleotidyl transferase -mediated dUTP nick end labeling(TUNEL) method and calculate apoptosis index(AI). Results:1. The change in general conditionsThe rats began to show the change in activity amount, appetite, body weight, clothing hair, stool 8 weeks after the models were made. The above-mentioned change was far more obvious by the end of the 24th week. The rats in the model group were emaciating, less active, shineless in hairs, poor in appetite with pasty and watery stool; The rats in the high-dose , middle-dose ,low-dose JWZJW groups appeared to improve to different degree . As far as the above-mentioned change was concerned, the rats of treti...