Experimental Research On The Effect Of HuaZhuoJieDuHeWei Decoction On The PCNA And PTEN In Rats Model Of Gastic Precancerous Lesions

Objective:By observing the impact of HuaZhuoJieDuHeWei Decoction on Gastic Precancerous Lesions model rats'general situation,gastric mucosal pathological histology , proliferating cell nuclear antigen(PCNA)and phosphatase tumor suppressor gene(PTEN), investigate the probable mechanism of HuaZhuoJieDuHeWei Decoction on the prevention and treatment of Gastic Precancerous Lesions and lay the foundation for further study of HuaZhuoJieDuHeWei Decoction.Method:80 SPF level healthy 6-week-old Wistar male rats randomly divided into 2 groups:normal-control group with 12 rats,model group with 68 rats.Except the normal-control group,the left were modeled by integrated modeling method for 24 weeks. After the model been successfully built,randomly divided the remaining 56 rats into 5 groups:the model group with 12 rats,the RetinoicAcid group, HuaZhuoJieDuHeWei Decoction high dose group, HuaZhuoJieDuHeWei Decoction middle dose group and HuaZhuoJieDuHeWei Decoction low dose group,each group with 11 rats. Begin to treat:the high dose group use high density HuaZhuoJieDuHeWei Decoction (2g crude drug/ml)to irrigate rats'stomaches according to 22g/kg·d.Similarly, the middle dose group(1g crude drug/ml)according to 11g/kg·d. The low dose group(0.5g crude drug/ml) according to 5.5g/kg·d. The RetinoicAcid group use retinoic acid liquid(density 4%)according to 40ml/kg·d. The corresponding amount saline was given to normal-control group and model group.All of the 6 groups were irrigated once a day for 16 weeks.After treatment, the rats were inhibited from eating and drinking water for 24 hours after last administration. Being weighed and anesthetized, all the rats were killed to take the stomaches.Then observe the pathology of the gastric mucosal tissue by eye and light microscope. Detect PTEN and PCNA in the gastric mucosal tissue through immunohistochemical methods. Measurement data are expressed with(±s).Use SPSS13.0 statistical software for statistical analysis.P<0.05 means the differengce is statistically significant.Result: (1)General condition of the rats:All the group of treatment can improve rats'fur luster,the state of mind,the appetite,the stool,the activity,the body weight and the survival rate. But the high dose group shows obvious influence. (2)Observe the gastric mucosa by eyes:The gastric mucosa surface of normal-control group are smooth and pink.The mucosal folds are regular. The mucous of the model group are crude,pale and with point sheet bleeds.Part of the plica are low or even disappear.Besides,erosion,ulcers and hyperplastic nodules are visible. The RetinoicAcid group and the TCM groups can improve the gastric mucosa pathological changes.The high dose group are close to the normal. (3)Gastric mucosa under light microscope observation:Gastric mucosa thickness of normal-control group is normal.The epithelial cell is complete.The glands are arranged neatly.The inflammatory cells are infiltrated occasionally.The gastric mucosa of model group multifocally becomes atrophy and thin.The epithelial cells have been degenerated,mortified and shedding.Mucous glands have different degrees of shrinkage or disappearance.Moderate dysplasia to severe dysplasia and intestinal metaplasia are visible.The muscularis mucosa becomes thicking. A large number of inflammatory cells are infiltrated in the interstitum.Gastric mucosa of high dose group and middle dose group are moderate thickness.The epithelial cells have no significant degeneration and necrosis.The glands are arranged regularly.A small number of inflammatory cells are infiltrated in the interstitum. Compared with the model group,all the groups of treatment have varying degrees of improvement in the gastric mucosal atrophy,intestinal metaplasia and dysplasia. But the middle dose group and the high dose group show obvious influence(P<0.05).(4)PCNA : The gastric mucosa of normal-control group express less positive cells and they are slightly stained. The model group express large number of positive cells and many of them are strongly positive with the color of dark brown. the TCM groups partly express less strong positive cells.PI of the model group is more than the other groups.PI of the high dose group is less than the RetinoicAcid group. The difference is significant(P<0.05).There is no significant differrnce between the middle dose group and RetinoicAcid group(P>0.05).The same to the low dose group.(5)PTEN:the gastric mucosa of normal-control group express large number of positive cells and they are stained dark brown.The average positive expression rate is 100%. The model group has the lowest average positive expression rate about 25.0%. There are significant differences between the model group and the other treatment groups(P<0.05).The positive expression rates of the high dose group and the middle dose group are higher than the other treatment groups.The differences are significant(P<0.05). (6)The correlation between the positive expression rate of PTEN and the PI value: r=-0.6032, P<0.05. Pointing that the correlation between the positive expression rate of PTEN and the PI value is negative. That is to say, accompany with the declining of the PTEN,the PCNA is increasing.Conclusion:(1) HuaZhuoJieDuHeWei Decoction can improve the general state of rat model of GPL.(2) HuaZhuoJieDuHeWei Decoction can improve the condition of gastritis,atrophy, metaplasia and dysplasia. It's efficacy has been authenticated further.(3)HuaZhuoJieDuHeWei Decoction may inhibit the excessive cell-proliferation through inhibiting the expression of PCNA and accelerating the expression of PTEN. Finally,it play the role of cure the GPL by rehabilitating the equilibrium of cell-proliferation and cell-apoptosis.