The Effect Of Weipixiao On Regulation On Glycolysis Signaling Pathway In Precancerous Gastric Leisions Rats

ObjectiveThe current study was designed to assess the pharmacological effects of weipixiao on gastric precancerous leisions(GPL)rats and the underlying molecular mechanisms.The expressions of LDHA,MCT4,CD147,HIF-1α,PI3 K,AKT,m TOR and mi RNA-34 a were examined.MethodsA total of 70 male Sprague-Dawley(SD)rats were randomly divided into control group which were administered routinely,the rest were model group providing with spontaneous intake of MNNG solution for 200mg/L and feeding every other day.At the 8th,12 th and 16 th week respectively,3rats of the model group were randomly sacrificed to perform histopathologic evaluation.Based on the histopathologic evaluation results,it was confirmed that the rats in model group were in the lesions of dysplasia.Then the model group were randomly divided into 4 groups: model group,positive group(Weifuchun,0.2g/kg),high dosage of WPX(WPX H)decoction(19.8g/kg)and low dosage of WPX(WPX L)decoction(9.9g/kg).10 weeks later,all rats were sacrificed.The gastric mucosa was divided into three parts,one was fixed with 10%neutral-buffered formalin to perform HE staining,HID-AB-PAS staining and immunohistochemistry,the second was fixed with 2.5% glutaraldehyde to observe the ultrastructure of the gastric epithelial cells,the third was saved in refrigerator which was used to assess the expressions of LDHA,MCT4,CD147,HIF-1α,PI3 K,AKT,m TOR and mi RNA-34 a.Results1.General state of the ratsIn control group,rats were in good spirit and nutriture,they were with burnished fur.when compared with the rats in control group,the rats in model group were in poor spirit with dull fur,they had lighter weight and fewer activities.When compared with the rats in modelgroup,rats in bose dosages of weipixiao had better spirit,more activities and heavier weight.These results indicated that weipixiao can improve the spiprt and diet of GPL rats.2.Histological analysisThe gastric mucosa and basilar membrane were in uniform thickness and gastric cavity was consisted of simple columnar epithelium in control group.Compared with control group,it could be observed that the thickness of basement membrane was asymmetrical in model group,and gastric mucosa was thinner,basilar membrane was thicker.In addition,dysplastic gastric epithelial cells with enlarged,hyperchromatic and crowded nuclei were distributed in mucoderm.Differentiated size of epithelial cells and cavity fusion of gastric mucosa was exited.All the results implied that dysplasia with intestinal metaplasia had presented in gastric mucosa.Interestingly,in both dosages of WPX decocion groups,the area of dysplastic gastric epithelial cells was decreased,indicating WPX decocion could ameliorate dysplasia of gastric mucosa in GPL rats.3.HID-AB-PAS stainingAdditionally,HID-AB-PAS staining was used to evaluate the degree of intestinal metaplasia.In control group,HID-AB-PAS staining was only limited to gastric cavity side,which may be caused by duodenal juice reflux.By contrast,intestinal metaplasia cells were situated in gastric cavity side and lamina proprian in model group.However,in both dosages of weipixiao groups,areas of two subtypes of intestinal metaplasia were attenuated,indicating that WPX decoction reversed MNNG-induced GPL.4.Ultrastructure analysisIn model group,dysplastic cells were clearly observed.The electronic density of cytoplasm were obviously reduced.The area of cytoplasm shrank leading to a dereased nucleus/ cytoplasm ratio.There was a reduction of the numbers of dysplastic cells in positive group.More importantly,in both doses of WPX decoction group,cells were main in vacuolation,and dysplastic cells were rare were rarely found.5.Expressions of glycolysis relative moleculesAccording to the results of RT-qPCR,the m RNA expressions of LDHA,MCT4,CD147,HIF-1α,PI3 K,AKT and m TORC1 were significantly increased in model group when compared with control group(P<0.01),while the level of mi RNA-34 a was markedly decreased(P<0.05).Additionally,the m RNA levels of LDHA,MCT4,CD147,HIF-1α,PI3 K,AKT and m TORC1 were significantly decreased in both doses of weipixiao groups(P<0.01,P<0.05),the expression of mi RNA-34 a was markedly increased(P<0.01).Manifested by the western blot results,the protein levels of LDHA,MCT4,CD147,HIF-1α,PI3 K,AKT and m TOR were significantly increased in model group(P<0.01),and both doses of weipixiao could down-regulate all the molecule expressions.Furthermore,consistent with the results of western blot,it could be observed that the protein levels of LDHA and m TOR were significantly increased in model group(P<0.01),and weipixiao could markedly down-regulate their levels(P<0.01).ConclusionsIn this study,the GPL rats were successfully established.the present study indicates that weipixiao reverse MNNG-induced GPL via regulating mi RNA-34a/ LDHA signaling pathway and mi RNA-34 a /PI3K/Akt /m TOR signaling pathway.Meanwhile it can also suppress glycolic process via restoring aberrance of MCT4,CD147 and HIF-1α.These findings may have important implications for the prevention of gastric cancer.