The Expression Of Notch1 In CD4+T Cell Of Peripheral Blood Of H.pylori-infected Patients And Its Significance

Background:Long-term colonization of gastric mucosa by Helicobacter pylori(H.pylori)can cause common gastric diseases such as gastritis,duodenal ulcer,gastric ulcer,gastric cancer and so on.In addition,it had been verify that there was a connection between HK pylori infection and ulcerative colitis.CD4+T cells mainly differentiate into Thl cells that secrete IFN-gamma to resist pathogen infection during H.Pylori-infected.The specific transcription factor T－bet gene is the key factor of Thl cell differentiation.which also promots the secretion of IFN-gamma factor.Recent studies had shown that Notch1 plays an important regulatory role in the polarization of CD4+T cells to Th1 cells what had been also demonstrated in some infectious diseases.Therefore,we speculated that Notchl receptor signal was involved in the differentiation of CD4+T cells into Thl cells by H.pylori-infeeted induction,which may be different in different parts of gastrointestinal diseases(gastritis,duodenal ulcer,gastric ulcer and ulcerative colitis).In this study,to evaluate whether Notchl is involved in Thl cell differentiation during H.pylori infection,and whether the Notch signaling pathway was activated,the expressions of Notchl,Hesl(the downstream target gene of Notch1),T-bet and IFN-gamma in CD4+T cells of H.pylori-infected patients will be detected.and the concentration of IFN-gamma in plasma will also be determined.Then,the correlation of Notchl with T-bet and IFN-gamma will be analyzed.At last.the role of Notch1 in different parts of gastrointestinal diseases with H.pylori infection,was evaluated.Method:The health people and the patients who receive treatment in the department of gastroenterology people’s hospital jiangmen city of xinhui district,from April 2018 to December 2018 were selected.Rapid urease assay(dipstick method)and 14C urea breath test were used to confirm H.pylori infection.And then,different parts of gastrointestinal diseases will be classifyied according to the results of electronic gastroenteroscopy and gastrointestinal mucosal biopsy in H.pylori-inmieeted patiens.Clinical and laboratory examination data were collected and analyzed.The mRNA expression levels of Notchl,Hesl,T-bet and IFN-gamma in CD4+T cells in peripheral blood of each group were detected by real-time quantitative fluorescence RT-QPCR at the same time.Western blot was used to detect the protein expression levels of Notch1 and Hesl in CD4-,T cells in peripheral blood of each group.Then,the expression of Notchl in HK pylori infection and inflanmmation in different parts deases will be analyzed at the same time.The correlation and significance of Notchl signal with T-bet and IFN-gamma were analyzed.Result:(1)72 patients who met the criteria were included in this study,among which 37 cases were confirmed positive for H.pylori infection,including 26 males(70%)and 11 females(30%)and the average age was(58.95±2.153)years.There were 35 healthy subjects,including 21 males,accounting for 60%,and 14 females,accounting for 40%and the average age of(53.63±2.584)years.There was no significant difference in gender and age between the two groups.Among the 37 H.pylori-infected patients,there were 18 cases of gastritis,5 cases of duodenal ulcer,5 cases of gastritis with duodenal ulcer,and 9 cases of ulcerative colitis,involving four different inflammatory diseases in different parts.(2)By comparing the results of the collected clinical data of blood routine inflammatory index test,the absolute numbers of leukocytes,granulocytes and monocytes of the two groups showed no statistical significance.The absolute number of lymphocytes of H.pylori-infected patients was lower than that in the healthy subjects,and the difference was statistically significant(P<0.01).The different parts of gastrointestinal diseases with H.pylori infection(gastritis,duodenal ulcer,gastric ulcer,ulcerative colitis)were analysis,the results showed that only the absolute number of lymphocytes in the gastritis group had statistic difference from the other three groups.(3)The mRNA expressions and protein levels of Notchl and Hesl in CD4+T cells of H.pylori-infeeted patients were higher than those in healthy subjects(p<0.0001、P<0.01).The mRNA expressions of T-bet and IFN-gamma in CD4+T cells of H.pylori-infected patients were higher than those in healthy subjects(P<0.05,P<0.05).Compared with the group only showing upper gastrointestinal inflammation,the mRNA expression levels of Notchl,T-bet and IFN-gamma were all increased in H.pylori-infected accompany the ulcerative eolitis(P＝0.001、P=0.03、P=0.02).However,there was no significant difference in the expression of Hesl mRNA between them(P=0.36).(4)Pearson correlation analysis showed that the mRNA expressions of T-bet and IFN-gamma were positive correlation with the mRNA expressions of Notchl in CD4+T cells of H.pylori-infected patients(r＝0.56,p<0.0001;r=0.41,p<0.05).However,there was no direct correlation between the concentration of IFN-gamma in plasma and the mRNA expressions of Notchl(r=0.02,P>0.05).Conclusion:The mRNA and the protein expression of Notchl receptor in CD4+T cells of H.pylori infection was increased.Moreover,the mRNA expression leves was different in different parts inflammatory diseases,it was obvious rised especially in ulcerative colitis.And the mRNA expression of Notchl was positively correlated with the mRNA expression of T-bet and IFN-gamma.It was suggested that the Notchl receptor was positively correlated with process of the differentiation of CD4+T cells into Th1 cells induced by H.pylori infection,and the mechanism can be futher studied from this pathway.