Regulating Mechanism Of Weipixiao On The Hypoxiao Tolerance And Aerobic Glycolysis In Gastric Mucosa In Rats With Precancerous Lesions Of Gastric Cancer

BackgroundGastric cancer is one of the most common tumor of malignance, which posses a high morbidity. A large number of epidemiological studies have shown that the precancerous lesion of chronic atrophic gastritis is associated with the occurrence of gastric cancer. Blocking and improving the gastric precancerous lesions is vital to reduce the incidence of gastric cancer and to prevent and treat gastric cancer. The mechanism of which the PLGC cancerates to gastric cancer is not clear at present, Western medicine only treats symptoms while there is definite curative effect in traditional Chinese medicine which can reverse PLGC and it is a hot spot in current research. So to clarify the pathogenesis of PLGC and explorethe medicine that can block and reverse the PLGC is critical.Objective1. To observe the effect of Weipixiao on the pathological organization structure and the ultrastructure of epithelial cells in Gastric Mucosa in Rats with Precancerous Lesions of Gastric Cancer.2. To observe the effect of Weipixiao on the expression of HK2> PKM2and LDH in Gastric Mucosa in Rats with Precancerous Lesions of Gastric Cancer.3. To observe the effect of Weipixiao on the microvascular ultrastructure and the expression of HIF-1a mRNAn HIF-1a protein and VEGF in Gastric Mucosa in Rats with Precancerous Lesions of Gastric Cancer.MethodsThe rats were randomized into six groups, including normal, model, Vatacoenayme and three groups of Weipixiao in high dose (15g/kg/d), medium dose (7.5g/kg/d) and low dose ((3.75g/kg/d). To prepare the rat model of PLGC by giving MNNG (200μg/ml) and basal diet, Xiaochengqi Decoction (2ml per) by gavage and abnormal of starvation and full which lasted18weeks. Dose groups would be given continuous medication from the ninth week to the eighteenth week. At the end of the eighteenth week, killed all the rats. And then to observe the ultrastructure of gastric mucosa microvessel and epithelial cells under the transmission electron microscope; to observe the pathological organization structure under the optical microscope; to measure the expression of HIF-1α mRNA in Gastric Mucosa with the method of RT-PCR; to measure the expression of HIF-1a protein and VEGF in Gastric Mucosa with the method of envision-typed immunohistochemistry; to measure the expression of HK2、PKM2and LDH in Gastric Mucosa.ResultsRats were successfully caused with PLGC. rats in normal group owned burnished fur, good spirit, flexibility, normal diet; Model group rats menifested messy and dull fur, spiritual malaise, being lazy to move, diarrhea which showed the typical spleen-deficiency syndron; WeiMeiSu group and the high, medium and low dose group of weipixiao shared dull fur, poor spirit, less motion and poor appetite to some extent r. But all of them were improved compared with the model group.Under the optical microscope, normal rats gastric mucosal cells were neat, which were made up of simple columnar epithelium and lamina propria. The nuclei of model group rats were hyperchromatic, hypertrophic, special-shaped, active-hyperplasiaed while glands were congested, irregular-morphologied, multiple layered, back-to-back, and among the stroma there were small lymphocyte infiltration and hyperplasia. The pathological organization structure of gastric mucosa were improved to varying degrees in high dose group, medium dose group and low dose group whereas it owned the best effect in the low dose group.Under the transmission electron microscope, in the model group, endothelial swelled heavily, Vascular lumen were severely narrow even blocked, basement membrane were rough and fractured, discontinuous, pericyte swelled or degenerated even disappeared. The ultrastructure of gastric mucosa microvessel were improved to varying degrees in high dose group, medium dose group and low dose group whereas it owned the best effect in the low dose group.Compared with the blank group, the expression of HIF-1a protein and VEGF in the model group expressed abnormally positive and there was a increasing tendency of the expression of HIF-1α mRNA; Compared with the model group, the Weipixiao decreased the expression of HIF-1α mRNA、HIF-1α protein and VEGF.Compared with the blank group, the expression of HK2and LDH in the model group decreased(P<0.05) and there was a diminishing tendency of the expression of PKM2;(2)Compared with the model group, the Weipixiao promoted the expression of HK2and increase the tendengcy of the expression of PKM2and LDH.Conclusion1. Pathological feature of animal models:the model group rats behaved as the obvious spleen deficiency syndrome; in pathological organization structure the mucosa epithelium menifested intestinal metaplasia and dysplasia; Also in ultrastructure it showed altered atypia such as nuclear pyknosis、 cytoplasm vacuoles and so on. It meaned that we duplicated the PLGC model with spleen deficiency successfully. The Chinese herbal compound weipixiao which invigorated spleen、removed blood stasis and detoxified could effectively improve spleen deficiency symptoms, reverse gastric mucosal intestinal metaplasia and dysplasia and pathological changes of epithelium ultrastructure to prevent PLGC to becoming gastric cancer2. Aerobic glycolysis characteristics:aerobic glycolysis and oxidative phosphorylation existed in gastric mucosa epithelial cells in PLGC rats simultaneously. Activity of HK2、PKM2and LDH decreased.Weipixiao might reverse sugar metabolic disorder through repairing the gastric mucosal injury.3. Etiology and pathogenesis of aerobic glycolysis:The mechanism of aerobic glycolysis in PLGC might be related to the injury of gastric mucosa microvascular and consequencely expression increasing of HIF-1α and VEGF. Microvascular injury induced hypoxic-ischemic damage in the gastric mucosa cells, secondarily increased expression of VEGF could not satisfy the needs of the normal gastric mucosa epithelial cells and damage repair. To adapt to hypoxic microenvironment, it resulted in aerobic glycolysis sugar metabolic disorder under the induction of HIF-1α.4. Interference effect of Weipixiao:Weipixiao might intervene hypoxia tolerance and aerobic glycolysis in gastric mucosa epithelial cells of PLGC rats through the repairing damaged blood vessels, reduce expression of HIF-1a and the VEGF...