Mechanism Of The Three Blood Treatment Of Rats With Chronic Atrophic Gastritis And Precancerous Lesions

Objective:Gastric precancerous lesions is the potential benign lesion of gastric carcinoma. Chronic atrophic gastritis(CAG) and intestinal metaplasia (IM) or dysplasia (Dy) due to CAG are the commonest of gastric precancerous lesions. We study the feature of the pathologic picture of precancerous lesion of CAG rats replicated by methods of pylorus spring insertion and hot paste by gavage. And we observe the influence of the therapy of Supplementing Qi and Activating Blood on morphological changes of gastric mucosa in rats with precancerous lesion of CAG. Further more, we lucubrate pathogenesy of precancerous lesion of CAG rats and the pharmaco-mechanism of precancerous lesion of CAG in rats treated with Supplementing Qi and Activating Blood of TCM at apoptosis and gene level.Methods: We replicate the rat model of precancerous lesion of CAG by methods of pylorus spring insertion and hot paste by gavage .We analysis the feature of the early apoptosis of precancerous lesion of CAG with Annexin V FITC/PI double labeled flow cytometry. The cDNA microarray consisting of 11060 clones of rat genes was used to detect and screen the differentially expressed genes in precancerous lesion of CAG rats and normal rats. The models of rats were randomly divided into five groups: the normal control group , spontaneous recovery group, Activating the blood and resolve blood stasis group, Regulate Qi and Activating the blood group, Supplementing Qi and Activating blood group. After treated with the therapies of Supplementing Qi and Activating blood , Activating the blood and resolve blood stasis and Regulate Qi and Activating the blood, we observe their influence on morphological changes of gastric mucosa in rats with precancerous lesion of CAG. At the same time we analysis all groups' apoptosis of precancerous lesion of CAG with Annexin V FITC/PI double labeled flow cytometry. One step method for extraction of total RNA of rats stomach tissue after administration by gavage of Supplementing Qi and Activating blood of TCM and the stomach tissue of the model group was used. After reverse transcription and fluorescent labeling by Cy3, Cy5, we get two groups of rats stomach cDNA probes and then hybridized with the cDNA gene expression profiles microarray hybridization. The result was scanned by the laser scanner and processed by software for image analysis, standardization, the ratio of value analysis, cluster analysis and Gene Ontology analysis . Results of gene array were authenticated by Real-Time PCR.Results:1 The model of precancerous lesion of CAG rats induced by pylorus spring insertion and hot paste by gavage was replicated. Pathologic picture show the rats model of precancerous lesion of CAG is convincing. 2 The ratio of the early apoptosis of the model control group increases more significantly than normal group (P<0.01).3 In the spontaneous recovery group the result of the light microscope showed that the gastric mucosa was atrophied and thinner, the glands was getting less, the various dysplasia were occurred. After the treating of the therapies of Supplementing Qi and Activating blood and Activating the blood and resolve blood stasis the inflammation of gastric mucosa and dysplasia were all relieved or disappeared.4 After the treatment the scores of pathomorphology in Supplementing Qi and Activating blood and Activating the blood and resolve blood stasis groups was significantly reduced than the spontaneous recovery group (P<0. 05 or P <0.01).5 The ratio of the early apoptosis of the model control group increases more significantly than the blank control group(P<0. 01). The ratio of the early apoptosis of the groups treated by three therapies of Activating the blood and resolve blood stasis decreases, Activating the blood and Resolve blood stasis and Regulate Qi and Activating the blood groups decrease more significantly than the model control group(P<0.01). Supplementing Qi and Activating blood group show the tendency of decreasing the ratio of the early apoptosis,but it has not the significant difference comparing with the model control group (P=0.057) .6 Totally 300 differentially expressed genes were identified in precancerous lesion of CAG rats in comparision with normal rats including 227 up-regulated genes and 73 down-regulated ones according to the designed data filter criteria. These differentially expressed genes belonged to many different functional gene families involving different biological processes.7 By analyzing the results of three chips, we found 205 differentially expressed genes, among which, there are 101 up-regulated expression genes and 104 down-regulated expression genes. These differentially expressed genes belonged to many different functional gene families involving different biological processes.8 Real-Time PCR authenticating results were consistent with those of micmarray study.Conclusion:1 The model of precancerous lesion of CAG rats induced by pylorus spring insertion and hot paste by gavage was replicated. Increasing the apoptosis may be one of the etiopathogenesis of precancerous lesion of CAG.2 The therapies of Supplementing Qi and Activating blood and Activating the blood and resolve blood stasis may have a better interrupt and reversal effect on the precancerous lesion of CAG, such as gastric mucosa atrophy and dysplasia.3 The therapies of Supplementing Qi and Activating blood, Activating the blood and resolve blood stasis and Regulate Qi and Activating the blood show inhibitory action on the early apoptosis of precancerous lesion of CAG rats. Restraining the apoptosis may be one of the effective mechanisms that the Activating the blood and resolve blood stasis on precancerous lesion of CAG rats reverse the gastric mucosa atrophy and dysplasia.4 A variety of genes involved in the formation of precancerous lesion of CAG rats. The 300 differentially expressed genes comprise the differential gene expression profile of precancerous lesion of CAG rats and describe the general changes in the gene expressions in precancerous lesion of CAG rats at transcriptional level. Further analysis of the identified genes might help reveal the molecular mechanism of precancerous lesion of CAG rats.5 These differentially expressed genes belonged to many different functional gene families involving different biological processes and gene regulations. Further analysis of the differentially expressed genes might help reveal the pharmaco-mechanism of Supplementing Qi and Activating blood of TCM at genic level.