| Objective: To assess early changes in serum histone H3 concentration in patients with urosepsis and its predictive ability for the onset of urosepsis.Exploring the regulatory mechanism of histone H3 promoting the onset of urosepsis.Methods: This study was divided into clinical experiment and basic experiment.In clinical experiment,a total of 80 patients who underwent percutaneous nephrolithotripsy were enrolled in the study and divided into control and urosepsis groups based on their postoperative outcomes.Serum histone H3 concentrations were detected using an enzyme linked immunosorbent assay,blood indexes were tested by automatic blood analyzers,vital signs data were obtained by monitors and manual measurements.These results were correlated with the incidence of postoperative urosepsis.Repeated measurements and receiver operating characteristic curves were employed to analyze early changes and predictive value of serum histone H3 concentration in urosepsis.A rat model of urosepsis was induced using ureteral ligation and lipopolysaccharide(LPS)injection method,modeling result was confirmed by immunohistochemistry.Serum histone H3 level was detected by enzyme linked immunosorbent assay(ELISA).The expression profiles of mi RNA and m RNA in sepsis rats were examined using mi RNA and m RNA sequencing and analyzed differentiated expression mi RNA and its downstream target genes.mi R-146b-5p and its target gene Paqr5 were selected for further investigation.Human Umbilical Vein Endothelial Cell(HUVEC)was treated with LPS to induce sepsis in vitro,histone H3 level was measured by ELISA and matrigel tube formation assay were performed to study the influence of LPS and histone H3 in vitro.mi R-146b-5p mimics and inhibitor were transferred into HUVECs,targeted regulating effect of mi R-146b-5p and Paqr5 was assessed by luciferase assay,quantitative real-time PCR(q PCR)and western blot(WB),and Paqr5 expression levels were quantitatively by immunofluorescence staining.Results: Sixteen of the 80 patients(20%)developed urosepsis after surgery.Our data showed significant intra-group differences in terms of postoperative histone H3 concentrations(P<0.0001)and variation trends(P<0.0001).Among analyzed blood markers,serum histone H3 concentrations 3 h post-operation(0.825 [95% confidence interval: 0.718–0.931,P<0.0001;cut-off value 256.74ng/ml,93.8% sensitivity,67.2% specificity])and 6 h post-operation(0.834 [95%CI: 0.721–0.947,P<0.0001,cut-off value 300.875 ng/ml,68.8% sensitivity,87.5%specificity])displayed a higher area under the corresponding receiver operating characteristic curves,indicating that these markers had a decent predictive value for postoperative urosepsis.Basic experiment showed that histone H3 levels were significantly increased in rats and HUVEC sepsis induced by LPS and triggered gene expression profile changes.mi RNA and m RNA sequencing displayed that,compared with the shamsurgery control group,14 mi RNA and 1132 m RNA were significantly upregulated,1 mi RNA and 949 m RNA were significantly downregulated.Luciferase assay,q PCR and WB verified the targeted inhibition of mi R-146b-5p on Paqr5.Mi R-146b-5p inhibitor could reverse the inhibitory effects of LPS and histone H3 on Paqr5.Conclusion: Our study suggests that serum histone H3 concentration is a novel predictor of postoperative urosepsis in patients undergoing percutaneous nephrolithotripsy.The findings of this study can be validated in a larger cohort.In vivo and in vitro experiments indicated that LPS could significantly increase the histone H3 levels both in vivo and in vitro.Histone H3 subsequently triggers differentiated expression of mi RNAs and m RNAs and promotes the onset of urosepsis targeted inhibiting Paqr5 through upregulation of mi R-146b-5p.Clinical Trial Registry Number:ChiCTR1800016679... |
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