| Objective: The model of chronic gastric atrophic gastritis precancerous lesions(PLGC)was established by using Kunming mice according to the comprehensive modeling method(MNNG load of ranitidine).The general condition of Yiwei Huayu Decoction and the pathological morphology of gastric mucosa were observed.The changes and the expression of STAT3,VEGF,CD31 protein and gene transcription were detected by immunohistochemistry and PCR,and the molecular biological mechanism of PLGC treatment was discussed.At the same time,the acute toxicity test of Yiwei Huayu Decoction was carried out to explore the safety of Yiwei Huayu Decoction.Methods: 1.Sixty SPF-class KM mice were divided into blank group 9 and model group 51 according to random number table method.In the blank group,0.9% sodium chloride injection was administered daily at 0.1 ml/10 g until the end of the experiment.After the model was molded by the comprehensive modeling method for 12 weeks,2 rats were randomly sacrificed,and the mold was confirmed by light microscopy.The remaining 45 models were divided into 5 groups according to the random number table method,which were model group,folic acid group,Yiwei Huayu decoction high-dose group,Yiwei Huayu decoction medium dose group,Yiwei Huayu soup small In the dose group,0.9% sodium chloride injection,0.002g/Kg folic acid suspension,20.54g/Kg Chinese medicine,10.27g/Kg Chinese medicine,5.14g/Kg Chinese medicine were administered respectively,and were administered once daily at 0.1ml/10 g.For 8 consecutive weeks.After fasting for 24 hours at the end of the 20 th week,the pathological changes of gastric mucosa were observed.The expression of STAT3,VEGF and CD31 protein in gastric mucosa was detected by immunohistochemistry,and the MVD value was calculated.The stomach was detected by real-time fluorescent quantitative PCR.Transcription of STAT3,VEGF,and CD31 genes in the mucosa.2.Forty healthy Kunming mice,male and female,weighing 20±2g,were randomly divided into two groups,the drug-administered group and the blank control group,with 20 rats in each group.After fasting for 12 hours,the drug-administered group was given the maximum concentration(4g/ml)and the maximum volume(0.4ml/10g)of Yiwei Huayu Decoction twice a day for 8 hours;the blank control group was given an equal volume by gavage.Distilled water.The rats were routinely fed after taking the drug,and the condition of the mice 14 days after the administration was observed and recorded.Result: 1.The experiment of Yiwei Huayu Decoction on STAT3/VEGF signaling pathway in gastric mucosa of PLGC mice: 1.1 General situation of mice in each group: Four rats died in the model group,two in the model group and folic acid group,and one in the low dose group.The mice in the blank group had good spirit,smooth fur,flexible movement,light red ears,more food,fast weight gain and normal urine and stool;the mice in the model group had poor mental state,rough fur,lack of color,less activity,pale ears,less food,slow weight growth,sparse dry stool and normal urine;the mice in the folic acid group had normal spirit,less shiny fur and one degree of activity.Generally,the mice in the large and medium dosage groups of Chinese herbal medicine had better mental state,slightly rough fur,relatively sensitive activities,increased food intake and weight gain than those in the model group.The mice in the low dose group had poor mental state,rough fur,and liked to pile up.Occasionally,the mice in the low dose group had pale ears,slightly increased food intake and body weight,but the change was not obvious.1.2 HE light microscopic observation of mice in each group: In the blank group,gastric mucosal cells were arranged neatly,continuously and intact,with normal cell shape and size,and no obvious inflammatory cell invasion was observed.In model group,the epithelial cells of gastric mucosa atrophied and thinned,the number of glands decreased,the structure of glands was disordered,inflammatory cells infiltrated,lymphoid follicles formed,and goblet cells formed slightly intestinal metaplasia.Compared with the model group,the other treatment groups had less invasion of inflammatory cells,relatively regular gland structure,increasing number of glands,homogeneous cell morphology,and gradually improved gastric mucosal lesions,especially in the large dose group of traditional Chinese medicine.1.3 Immunohistochemistry was used to detect the expression of STAT3/VEGF pathwayrelated protein in gastric mucosa of PLGC mice.Compared with the blank group,the expression levels of STAT3 protein,VEGF protein and CD31-MVD in the model group were significantly increased(P<0.01).Compared with the model group,the expression of STAT3 protein in each drug treatment group decreased significantly,which was statistically significant(P<0.01).Compared with the model group,the expression of VEGF protein in each drug treatment group decreased significantly,which was statistically significant(P<0.01).Compared with the model group,the CD31-MVD value of the low-dose group of traditional Chinese medicine decreased statistically(P<0.05);the decrease of CD31-MVD value in the folic acid group,the high-dose group and the middle dose group was statistically significant(P<0.01).).1.4 The transcription of STAT3/VEGF pathway related genes in gastric mucosa of PLGC mice was detected by PCR.Compared with the blank group,the expression levels of STAT3 m RNA,VEGF mRNA and CD31 m RNA in the model group were significantly increased(P<0.01).Compared with the model group,the expression levels of STAT3 m RNA in the folic acid group and the high-dose and medium-dose groups of Chinese medicine were decreased,which was statistically significant(P<0.01).Compared with the model group,the expression of VEGF m RNA in the folic acid group and the high-dose group of traditional Chinese medicine decreased,which was statistically significant(P<0.01).Compared with the model group,the expression of CD31 m RNA in each treatment group decreased,which was statistically significant(P<0.01).2.Acute toxicological experiment of Yiwei Huayu Decoction: 2.1 General conditions of mice in each group: The mice in each group were in good general condition,with normal diet and drinking water,soft fur,sensitive reaction,free movement,no obvious abnormality in appearance,no stool formation,no black stool and so on.All mice in each group survived until the end of the experiment,and no death was found.2.2 There was no significant difference in the body weight of mice in acute toxicological test on the first day,the first day,the third day,the seventh day and the fourteenth day before the formal experiment.2.3 In acute toxicology experiment,there was no significant difference in organ index between the administration group and the blank group.2.4 The lethal rate(LD50)of mice in the Yiwei Huayu Decoction group was less than half,and the total dosage of the drug was 320g/Kg,which was 243 times of the clinical dosage.Conclusion: 1.In this experiment,the PLGC model was replicated by the comprehensive modeling method of MNNG load-filled ranitidine.After 12 weeks of modeling,the pathology under light microscopy confirmed the successful modeling.Yiwei Huayu Decoction can improve the general condition of mice and the pathological changes of gastric mucosa.At the same time,STAT3,VEGF and CD31 in the gastric mucosa of mice were significantly decreased after intervention with Yiwei Huayu Decoction,suggesting that Yiwei Huayu Decoction may improve the stomach by inhibiting the microangiogenesis of precancerous lesions and reducing the possibility of gastric cancer.The pathological state of the mucosa achieves the purpose of treating PLGC.2.Yiwei Huayu Decoction is safe and reliable. |
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