LncRNA CYP1B1-AS1 as a clinical biomarker exacerbates sepsis inflammatory response via targeting miR- 18a- 5p

Figures

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  Fig. 1. Elevated serum CYP1B1-AS1 has potential as a diagnostic biomarker of sepsis. A Differential expression volcano plots of lncRNA in sepsis from the GSE217700 database. B Serum CYP1B1-AS1 expression in the subjects. C-D Correlation analysis between serum CYP1B1-AS1 and severity scores (SOFA and APACHE II score). E ROC curves were employed to evaluate the diagnostic utility of CYP1B1-AS1 in sepsis patients

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  Fig. 2. CYP1B1-AS1 levels may predict unfavorable outcomes in sepsis patients. A Serum CYP1B1-AS1 expression in death and survival patients with sepsis at 28 days of follow-up. B The ROC curve was employed to examine the predictive significance of serum CYP1B1-AS1 for mortality in sepsis patients. C Kaplan-Meier curves depicted cumulative survival rates, with group differences assessed by the log-rank test. **** P < 0.0001

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  Fig. 3. Silencing CYP1B1-AS1 mitigates LPS-induced effects on THP- 1 cell proliferation, apoptosis, and inflammation. A Impact of LPS concentrations on CYP1B1-AS1 levels in THP- 1 cells after 48 h. B Impact of LPS induction (1 µg/ml) on CYP1B1-AS1 levels in THP-1 cells. C RT-qPCR assessment of CYP1B1-AS1 siRNA transfection efficiency in THP- 1 cells. D RT-qPCR analysis of CYP1B1-AS1 expression in THP- 1 cells treated with LPS and si-CYP1B1-AS1. E-F. Flow cytometry and ELISA assay were used to assess cell apoptosis and inflammation factors in LPS and si-CYP1B1-AS1-treated cells. ** P < 0.01, *** P < 0.001, **** P < 0.0001

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  Fig. 4. CYP1B1-AS1 directly interacts with miR- 18a- 5p. A Investigation of CYP1B1-AS1 subcellular localization in THP- 1 cells. B Venn diagram presented the overlapping target miRNAs of CYP1B1-AS1 predicted by LncBooK2 and DIANA databases. C The putative binding sequences were presented. The targeting relationship between CYP1B1-AS1 and miR- 18a- 5p was validated using Dual-luciferase report assay (D), RIP assay (E), and RNA pull-down assay (F). G The serum miR- 18a- 5p in the subjects was detected. H Pearson coefficient was conducted to examine the correlation between serum miR- 18a- 5p and CYP1B1-AS1 levels in patients with sepsis. I The mRNA levels of miR- 18a- 5p were altered in THP- 1 cells that were induced with LPS and transfected with si-CYP1B1-AS1. ** P < 0.01, *** P < 0.001, **** P < 0.0001

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  Fig. 5. miR- 18a- 5p downregulation partly reversed the effects of miR- 18a- 5p silencing on LPS-induced cell damage in THP- 1 cells. A Changes in miR- 18a- 5p levels in THP- 1 cells after 48 h of induction with various LPS concentrations. B miR- 18a- 5p levels vary in THP- 1 cells induced by 1 µg/ml LPS for different durations. Transfection with miR- 18a- 5p inhibitor alone (C) or LPS-induced silencing of CYP1B1-AS1 alters THP- 1 cells (D). E-F Flow cytometry and ELISA assay were used to assess cell apoptosis, and inflammation factors in LPS, si-CYP1B1-AS1, and miR- 18a- 5p inhibitor CO-treated cells. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001