Helicobacter Pylori And Gastric Mucosal Lesions In The Relationship Between Basic And Clinical Research

Objective: To investigate the role of SHP2in the process of STAT3regulating IL-6by Helicobacter pylori LPS.Methods: Hp-LPS with the concentrations of0μg/ml,1μg/ml,10μg/ml affectedgastric cancer cell for12hours. The protein levels of STAT3and SHP2were detectedby western blotting, while the mRNA level was determined by Real time PCR. Weobserved that Hp-LPS with the concentrations of1μg/ml affected gastric cancer cellin different points such as0minute,1minute,10minutes,30minutes and60minutes.We detected the protein levels of P-STAT3and P-SHP2by western blotting. Hp-LPSwith the concentrations of0,1μg/ml,10μg/ml,20μg/ml,40μg/ml affected gastriccancer cell for30minutes. The protein levels of P-STAT3and P-SHP2were detectedby western blotting. After NSC87877with the concentration of60μg/ml inhibittingSHP2for six hours, we interfered with gastric cancer cell with Hp-LPS by theconcentration of1μg/ml. We detected the levels of P-STAT3, P-SHP2and P-ERKbetween treated group by NSC87877and untreated group. After SHP2inhibitted byNSC87877with the concentrations of60μg/ml for six hours and gastric cancer cellinterfered with by Hp-LPS with the concentrations of1μg/ml. We detected the levelsof P-STAT3and P-ERK by immunofluorescence cytochemistry staining techniques.And IL-6was detected by ELISA.Results: Helicobacter pylori LPS with the concentrations of0μg/ml,1μg/ml,10μg/ml affected gastric cancer cell for12hours. The protein level of STAT3and SHP2had no significant difference, when P value was larger than0.05. The result of Realtime PCR showed that mRNA level of STAT3increased with the concentration ofLPS increasing. However, that of SHP2had reverse result. We also interfered withgastric cancer cell in different points such as0minute,1minitue,10minitues,30minitues and60minutes. The level of P-STAT3was highest at30minutes by westernblotting, when P value was smaller than0.05. But P-SHP2had not display the same result. We interfered with gastric cancer cell in different concentrations such as0μg/ml,1μg/ml,10μg/ml,20μg/ml and40μg/ml. Datas showed that P-STAT3had aconcentration dependence, while P-SHP2had no concentration dependence. Weinhibitted SHP2by NSC87877with the concentration of60μg/ml for six hours. Afterthat we interfered with gastric cancer cell with LPS with the concentration of1μg/ml.We detected the levels of P-STAT3, P-SHP2and P-ERK between treated group byNSC87877and untreated group. We found that the level of P-STAT3was higher intreated group than that of untreated group, while those of P-SHP2and P-ERK werelower. We also found the level of P-STAT3was higher than that of untreated groupand the level of P-ERK was lower than that of untreated group byimmunofluorescence cytochemistry staining techniques. We found that the level ofIL-6was higher than that of untreated group by ELISA.Conclusion: In invitro study, the result displayed that Helicobacter pylori LPS couldactivate STAT3and induce the expression of IL-6. When SHP2was inhibitted byNSC87877, the level of IL-6promptly increased than that of untreated group. SHP2was an important regulator, which played an important role in the process ofHelicobacter pylori LPS activating STAT3and the expression of IL-6. Objective: To investigate the effects of Tip-α on cytokines expression. Whether theeffects depend on the signal way of NF-κB.Methods: We thus cloned Tip-α from genome sequence of H. pylori strain26695.The recombinant plasmid was transformed into E.coli. The expression product wasconfirmed by western blotting. We interfered with two kinds of cells in different timesby Tip-α in the concentration of12.5μg/ml and we detected the concentrations ofcytokines by ELISA. We also interfered with cells by different concentrations ofTip-α, we detected the state of cytokines by ELISA. After NF-κB inhibitted by PDTC,we interfered with two kinds of cells by Tip-α in the concentration of12.5μg/ml, wedetected the state of cytokines by ELISA.Results: SDS-PAGE showed a23ku protein identified by western blotting analysis.When we interfered with the cells by Tip-α in the concentration of12.5μg/ml indifferent times, the levels of cytokines such as IL-1β, IL-8, TNF-α were higher thanthat of without interferring with the cells. When we interfered with the cells by Tip-αof12.5μg/ml for two hours, the levels of cytokines were most high. But datas did notshow evident time dependence. When we interfered with the cells by differentconcentrations of Tip-α for two hours, the levels of cytokines were higher than that ofwithout interferring with the cells. The levels of cytokines by Tip-α of12.5μg/ml fortwo hours were most high. But datas did not show evident concentration dependence.In addition, the levels of cytokines of SGC-7901was higher than that of GES-1byTip-α of12.5μg/ml for two hours. When NF-κB inhibitted by PDTC, we found thatthe levels of cytokines significantly decreased than non-inhibitted by PDTC.Conclusion: Tip-α may play an important role on the expression of cytokines byNF-κB. Objective: To investigate the effect of folic acid combined with helicobacter pylori(Hp) eradication therapy on Atrophic Gastritis.Methods: From December2009and March2011at the First Affiliated Hospital ofNanjing Medical University,184gastroscopy and pathological examinationdiagnosed chronic Atrophic Gastritis patients were selected. Of those,90cases wereHp positive and94cases were negative. H. pylori positive patients were randomlydevided into group a and group b,43patients in group a were treated with standardtriple H. pylori eradication therapy and followed up3months of folic acid therapy.47patients in group b and Hp negative patients were with three months of folic acidtherapy. The clinical symptoms of each group was scored at before treatment, onemonth folic acid therapy and three months folic acid therapy and analyzed by singlefactor analysis of variance and t test. Patients of each group received gastroscopyexamination at before treatment and3months after medicine withdrawal, endoscopicscores, pathological scores and t test were recorded. The serum levels of pepsinogen(I, II) and gastrin17in venous blood of each group were detected by ELISA methodat before treatment and3months after medicine withdrawal.Results: Compared with before treatment, after one month folic acid therapy, therewas no statistical significance in clinical symptoms score of all patients(p=0.17,t=1.38). There was statistically significant compared with3month therapy. By theend of therapy, clinical symptoms score of group a was lower than that of group b,and the difference was significant(t=6.00, P=0.00). There was statistical significancein endoscopic scores of all patients between before treatment and aftertreatment(t=11.12, P=0.00). There was no statistical significance in endoscopic scoresbetween group a and group b after the treatment. The differences in each pathologicalscore of all patients (inflammatory scoring, active scoring, atrophy scoring,intestinal metaplasia scoring, atypical hyperplasia degree scoring) were significant between before treatment and after treatment. The level of pepsinogen I beforetreatment was lower than that of after treatment (t=3.19, P=0.00). There was nosignificant difference in the level of pepsinogen II before treatment and aftertreatment(t=0.25, P=0.81). After treatment the level of gastrin17was higher than thatof before treatment(t=5.33, P=0.00).Conclusion: Folic acid in combination with helicobacter pylori eradication can befavorable of Atrophic Gastritis, which may promote the secretion of pepsinogen andgastrin.