| Background:In China,early gastric cancer detection has long been in a low rate,which lead to an unsatisfied result of gastric cancer treatment.Chronic atrophic gastritis(CAG)represents a key status during gastric cancer(GC)development,and as a precancerous gastric lesion,its treatment can effectively enhance the whole rate of GC prognosis.To date,no convincing treatment was available for reversal of atrophic gastritis.Our team has proved through a IV stage multicenter clinical prospective study that a compound containing whole extract from suckling lamb gastric mucosa and vitamin B12 can effectively reverse atrop hic and intestinal metaplasia lesions in CAG patients.To understand the underlying mechanisms,we introduce Wistar rats to carry out our following experiments.Aim:To confirm the reversal role of a compound containing whole extract from suckling lamb gastric mucosa and vitamin B12 for CAG and explore the underlying mechanisms involved.Methods: 1.To establish successful CAG lesion in rats,Wistar rats were administered 100?g/ml MNNG water resolution for daily drinking,standard murine feed containing0.03% renitidine was provided for food,in addition,rats in the experimental group also received a starvation treatment followed by a 50% alcohol gavage once a week for 20 weeks.Twenty weeks later,all rats were kept for normal diet for another 1.5 month,then some rats were sacrificed after starvation for 24 h,The gastric juice,serum and gastric tissue samples were collected from each group.The pathological changes of gastric mucosa were observed by HE staining,immunohistochemical staining and AB-PAS staining to assess the severity of the lesion,which is important for the next experiment stage of assessing the efficacy of the drug.Using the corresponding kit to detect the relevant serum oxidative stress factor to determine whether the occurrence of oxidative stress response happens.2.The rats left were divided into two groups,rats in the treatment group were administered the drugs according to a weight ratio of 0.02 / 100 g,and the rats in the control group were given the same amount of normal saline for 3 months.After starvation,the rats were sacrificed to gather certain specimens.Water and food intake and body weight changes and other data were recorded weekly during this period.HE staining ?Immunohistochemical staining and AB-PAS staining were used to detect the pathological changes of gastric mucosa in rats after treatment.3.By detecting oxidative stress factors including SOD?GSH-Px and MDA levels in rats serum and Ki-67 and Bax staining,we primarily detected the underlying mechanisms concerning the reversal of rat CAG lesions.Results:1.The establishment of chronic atrophic gastritis in rats: During the modeling period,the water intake amount and food intake amount of the rats in the model group were significantly lower than those in the normal control group,and the body weight change was statistically different from the 4th week 240.16 ± 21.62 g vs.normal group 291.00 ± 35.52 g,P <0.05).At the end of modeling,the p H value of gastric juice in model group was higher than that in normal control group(model group 3.56 ± 0.62 vs.normal group 2.01 ± 0.56,P <0.05).HE staining showed that the gland of the antral mucosa of the normal control group was abundant,the gland morphology was normal,no inflammatory cell infiltration,glandular atrophy or structural abnormality were observed.The gastric antrum of the model group the number of lamina propria glands is significantly reduced,glandular structure disorder,arranged to lose the rules,different sizes,glandular space widened,there is obvious inflammatory cell infiltration.The gastric lesions of the two groups were compared according to the standard scoring systems,and we found that inflammation(model group 2.30 ± 0.64 vs.normal 0.00 0.00 0.00,P <0.05),atrophy(model group 2.20 ± 0.63 vs.normal 0.00 0.00 P <0.05)and intestinal metaplasia(0.70 ± 0.82 vs.0.00 ± 0.00,P <0.05)lesions were all statistically significant.In addition,the mucosal layer of the model group was significantly thinner,the thickness of the mucosal muscle layer increased,the thickness ratio of mucous membrane and mucosal muscle layer decreased significantly(6.34 ± 1.05 vs.2.41 ± 0.77,P <0.05).AB-PAS staining showed that the density of gland in model group was significantly lower than that in model group(model group 7.53 ± 2.31 vs.normal group 13.43 ± 1.26,P <0.05).The results showed that the atrophy rate of the model group reached 100% and the intestinal rate was 50%.There was significant difference in atrophy and intestinal formation between the two groups.The levels of SOD,GSH-Px and MDA in the two groups were significantly lower than those in the normal group(P <0.05),and the levels of SOD and GSH-Px in the model group were significantly lower than those in the normal group(P <0.05;MDA 17.33 ± 7.59 vs.8.36 ± 4.82,P <0.05),and the concentration of SOD was significantly lower than that of the normal group(SOD vs 34.45 ± 15.43 vs.87.55 ± 12.21,P <0.05;GSH-Px 13.22 ± 8.37 vs.41.35 ± 5.46,P <0.05).2.After treatment period,it was found that the weight gain of the rats in the treatment group was gradually higher than that in the non-treatment group,and the differences in body weight between the two groups were statistically significant(508.26 ± 33.96 vs.treatment group 495.50 ± 23.01,P <0.05)from the sixth week.The p H value of the gastric juice in the treatment group was slightly higher than that in the normal control group(P <0.05),but lower than that in the non-treatment group,the difference was statistically significant compared with the untreated group(3.07 ± 0.55 vs non – treatment group,4.45 ± 0.72,P <0.05).Statistical analysis was performed on the lesion of the treatment group and the non-treatment group.No significant difference was found between the inflammation scores(treatment group 1.43 ± 0.53 vs.non-treatment group,2.14 ± 0.69,P = 0.051)but in the atrophic scores(treatment group 1.57 ± 0.79 vs.non-treatment group 2.71 ± 0.49,P = 0.007)and intestinal metaplasia lesion scores(treatment group 0.71 ± 1.50 vs.non-treatment group 3.57 ± 0.79,P = 0.001).The thickness of gastric mucosa and mucosal muscle in rats were similar to those in normal control group.The thickness of gastric mucosa was significantly decreased in non-treatment group and the thickness of mucosal muscle layer was significantly increased.The ratio in the non-treatment group was significantly higher than that of the treatment group(treatment group 3.78 ± 0.45 vs.non-treatment group,2.37 ± 0.38,P <0.05).The mean number of glands in the gastric mucosal unit length(1 mm)was 7.47 ± 2.06,which was significantly lower than that of the rats in the untreated group(9.58 ± 1.51,P <0.05).Statistical analysis of the number of gastric mucosal surface nodules found that in the corpus,antrum and whole stomach area,the numbers in treatment group and in the non-treatment group were statistically different(see Figure 2-3 in the second part of the text).The results showed that the severe atrophy rate and intestinal metaplasia rate of the rats in the treatment group were significantly lower than those in the non-treatment group(severe atrophy rate: 14.29% vs.non-treatment group,71.43%,P <0.05;intestinal rate: treatment group 28.58% vs.non-treatment group 57.14%,P <0.05).3.The underlying mechanisms of rats CAG reversal:Ki-67 staining showed that rats in the treatment group demonstrated higher proliferative scores than those in the normal control group,Bax staining showed a reduced apoptosis trend in the glands of the treatment group rats than those in the non-treatment group.In addition,SOD and GSH-Px activities were increased after receiving treatment of the capsules,whereas MDA level was decreased.Conclusions:1.Wistar rats given free access to 100?g/ml MNNG solution,0.03% of ranitidine feed,and 50% alcohol following 24 h starvation for 20 weeks developed atrophic gastritis successfully with high appearancing rate and high stable rate.2.The drug containing whole extract from suckling lamb gastric mucosa and vitamin B12 can successfully reverse atrophic gastritis lesions in rats.3.The mechanisms underlying the reversal of rat gastric atrophy by the drug containing whole extract from suckling lamb gastric mucosa and vitamin B12 may involve in the increase in normal gland formation and inhibition of oxidative stress reaction and reduced apoptosis in the mucosa gland layer. |
PDF Full Text Request