| Objectives:1.The results of several recent published studies have shown that thrombocytopenia improves in approximately half of cases after H.pylori(Hp)eradication in ITP patients,however,most of the studies included in the study were observational cases,we propose to use conventional and reticulated Meta-analysis based on randomized controlled trials in o rder to better understand the effects of Hp eradication in ITP patients and also to select the optimal therapy.2.To investigate the regulatory effect of Hp on Dami cell apoptosis through the in vitro action of H.pylori on human megakaryocyte Dami cells,and to explore how Hp regulates Dami cell apoptosis through the NF-κB signaling pathway and the specific molecular mechanism in combination with the NF-κB specific inhibitor phthalopyrrolidine dithiocarbamate(PDTC),in order to provide a theoretical basis for the clarification of how Hp acts on ITP in cytology.3.To investigate the therapeutic effects of PDTC,a specific inhibitor of NF-κB,on HP-infected ITP mice at the animal level and the possible mechanism of promoting platelet recovery in ITP mice,in order to provide new ideas and targets for clinical prevention and treatment of ITP.Methods:1.A systematic computer search of electronic databases including PubMed,EMBASE,Google Scholar and Cochrane Library)was conducted according to the set search strategy.Screening was performed according to the established inclusion and exclusion criteria,and data extraction and quality evaluation were also performed for the final included literature.The extracted data were analyzed for traditional Meta heterogeneity and direct comparison using Review Manager software,while mesh Meta analysis was performed using ADDIS software.2.Dami cells were treated with Hp in vitro at 0,3,6 and 9 h.The apoptosis rate of each group was assessed by flow cytometry,the morphological changes of each group were observed by transmission electron microscopy,and the expression of Bcl-2 and Bax related to apoptosis and inflammatory genes NF-κBp65,were assessed by fluorescence quantitative PCR and Western blot immunoblotting.IL-17 expression.After treatment of HP-infected Dami cells with PDTC for 6 h,the apoptosis rate,cell morphology of each group,expression of Bcl-2 and Bax,and expression of inflammatory genes NF-κB p65 and IL-17 were assessed in the same manner.3.Mice were randomly divided into Normal group,ITP group,SSI+ITP group and SSI+ITP+PDTC group.The SSI+ITP group and SSI+ITP+PDTC group were infected with HP by gavage with HP bacterial solution,and the ITP model was established by intraperitoneal injection of anti-mouse CD41 monoclonal antibody method,and the SSI+ITP+PDTC group was treated with simultaneous The mice were treated with PDTC by gavage.At the end of modeling,platelet counts were measured,mice were executed,gastric tissues were taken for pathological observation,femoral sections were taken for megakaryocyte counting and morphological observation,sternal bone marrow smears were taken to observe the morphology and primary classification of megakaryocytes under oil microscope,and flow cytometry was used to analyze the positivity of mature bone marrow megakaryocytes(CD41+CD42+),and thymus and spleen were weighed to calculate the organ coefficients.Results1.A total of 241 patients(125 in the erad ication group and 116 in the control group)were finally included in a meta-analysis of 6 randomized controlled studies.the total platelet response rate was significantly higher in the Hp eradication group than in the control group(risk ratio=1.93,95%confidence interval:1.01-3.71,P=0.05).In subgroup analysis,children in the Hp eradication group failed to show a statistically better response rate than the non-eradication group(hazard ratio=1.80,95%confidence interval:0.88-3.65,P=0.11).Among the six treatment modalities involved in this study,the best treatment modality for ITP was the hormone combined with Hp triple eradication strategy.2.After Hp treatment of Dami cells,there was a significant difference in the apoptosis rate of Dami cells between the groups,and the apoptosis rate gradually increased with the prolongation of Hp exposure time,with the most significant change at 6 hours and a significant difference between 0 and 6 hours.3.There were significant differences in the morphological changes of apoptotic cells between groups after Hp treatment of Dami cells.4.After Hp treatment of Dami cells,the expression of Bax,p-p65 and IL-17 gradually increased with the prolongation of HP exposure,and the expression of anti-apoptotic Bcl-2 protein gradually decreased,which was significantly different compared with the control group.5.After the intervention of Hp-treated Dami cells with NF-κB-specific inhibitor PDTC,although the apoptosis rate was higher than that of the control group,it was lower than that of the group treated with HP alone,while the apoptotic morphology showed some degree of improvement.6.After intervention with PDTC on Hp-treated Dami cells,the expression of Bax,p-p65,IL-17 and Bcl-2 showed significant reversal,and compared with the group treated with HP alone,the expression of Bax,p-p65 and IL-17 decreased and the expression of Bcl-2 increased,with significant differences.7.Hp infection could significantly increase the optical density value of anti-Hp antibody in peripheral blood and inflammatory infiltration and atrophy of gastric mucosal epithelium in mice.8.Compared with ITP model mice,the number of platelets in peripheral blood of HP infected ITP model mice continued to decrease,and the classification and morphology of bone marrow megakaryocytes changed abnormally.These changes were significantly improved after the intervention of PDTC.9.PDTC intervention treatment can increase the thymus index and reduce the spleen index of HP infected ITP model mice,which was significantly different compared with the model group and HP infection model group.10.Compared with the model group and HP infection model group,PDTC intervention treatment can significantly reduce the number of megakaryocytes and granular giant cells in the bone marrow of ITP mice,and increase the number of plate giant cells.11.The number of mature megakaryocyte lineage progenitors(CD41+CD42+)was reduced in the ITP model mice,and HP infection further reduced the number of mature megakaryocyte lineage progenitors(CD41+CD42+)in the bone marrow of mice,while PDTC intervention treatment promoted the expression of CD41 and CD42b.12.Compared with the control group,the number of TUNEL positive cells in ITP model group increased significantly,and NF-κB signal was significantly up-regulated.After HP bacteria infected ITP model,the number of TUNEL positive cells continued to increase and NF-κB signal is further up-regulated.However,HP bacteria infected ITP model mice treated with PDTC can significantly reduce the number of TUNEL positive cells and NF in bone marrow-κB signal activation.Conclusions1.Hp eradication has a significant therapeutic effect in patients with ITP;hormone combined with Hp triple eradication is the best ITP treatment;considering the inherent limitations of the sample size of the included studies,a large-scale randomized controlled trial is needed to validate the therapeutic effect of Hp eradication in adults and pediatric patients with ITP.2.H.pylori infection can significantly promote the apoptosis of megakaryocytes and aggravate the progression of ITP disease.Its mechanism may be related to NF-κIt is related to the activation of B/IL-17 pathway.That is,it mediates the apoptosis of ITP bone marrow megakaryocytes through NF-kB/IL-17 signal loop,inhibits the production of peripheral platelets,and aggravates the occurrence and development of ITP.3.Pyrrolidine dithiocarbamate(PDTC)can promote the recovery of platelet count in HP infected ITP model animals.Its mechanism may reduce the injury and apoptosis of bone marrow megakaryocytes to a certain extent,promote the recovery of bone marrow megakaryocyte function and slow down the occurrence and development of ITP by inhibiting the over activation of NF-kB.This may be the mechanism of pyrrolidine dithiocarbamate in the clinical treatment of ITP. |
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