Revealing the role of Peg13: A promising therapeutic target for mitigating inflammation in sepsis

Abstract

To investigate the role of Peg13 in modulating the inflammatory response in sepsis, we established Lipopolysaccharide (LPS)-induced 293T cells and mouse models. Peg13 expression was assessed at various time points after infection using RT-qPCR. The levels of high mobility group box 1 (HMGB1) and interleukin-6 (IL-6) were quantified through ELISA. A total of 44 septic patients and 36 healthy participants were recruited to measure Peg13 and HMGB1 levels in the blood. Peg13 demonstrated significant down-regulation in the supernatant of LPS-induced 293T cells and in the blood of LPS-induced mice. Moreover, the levels of proinflammatory cytokines HMGB1 and IL-6 were elevated in both the supernatant of LPS-induced cell models and blood specimens from LPS-induced murine models, and this elevation could be notably reduced by Peg13 suppression. In a clinical context, Peg13 and HMGB1 levels were higher in septic patients compared to healthy subjects. Peg13 exhibited a negative correlation with HMGB1, C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) among septic patients. Peg13 mitigates the inflammatory response by reducing the release of proinflammatory cytokines HMGB1 and IL-6 in sepsis, presenting a potential therapeutic target for alleviating inflammation in sepsis treatment.

Figure 1 -. Peg13 Up-regulation in LPS-induced 293T cell supernatant silenced by shRNA Lentivirus (sh-Peg13). (A) Fluorescent images of 293T cells infected with 10 μL sh-Peg13 lentivirus (MOI=10, 50 or 100) along with (M) 90 μL complete culture medium; or (A) 86 μL complete culture medium and 4 μL HiTrans G A; or (P) 86 μL complete culture medium and 4 μL HiTrans G P at 72 h. (B) Fluorescent images of 293T cells exposed to 50 ng/mL LPS and infected with sh-NC or sh-Peg13 lentivirus at 48, 72, and 96 h. (C) Peg13 expression in 293T cells under control, LPS, LPS + sh-NC lentivirus or LPS + sh-Peg13 lentivirus conditions at 16, 48, 72, and 96 h. **p<0.01; ***p<0.001; ****p<0.0001.

Figure 2 -. Peg13 suppression aggravates LPS-induced proinflammatory cytokines HMGB1 and IL-6 release in 293T Cells. (A-D) HMGB1 content in 293T cells treated with 50 ng/mL LPS and infected with sh-NC or sh-Peg13 lentivirus at 16, 48, 72, and 96 h. (E-H) IL-6 level in 293T cells exposed to 50 ng/mL LPS or/and infected with sh-NC or sh-Peg13 lentivirus at 16, 48, 72, and 96 h. *p<0.05; **p<0.01; ****p<0.0001.

Figure 3 -. Peg13 expression is up-regulated in septic mice blood and its decrease by sh-Peg13 Lentivirus. (A-C) Peg13 expression in the blood of control mice, LPS-induced septic mice, and septic mice injected with sh-NC or sh-Peg13 lentivirus at 3, 7, and 14 days. **p<0.01; ***p<0.001; ****p<0.0001.

Figure 4 -. Peg13 Inhibition Exacerbates Proinflammatory Cytokines HMGB1 and IL-6 in Septic Mice Blood. (A-C) HMGB1 content in blood of control mice, LPS-induced septic mice, and septic mice injected with sh-NC or sh-Peg13 lentivirus at 3, 7, and 14 days. (D-F) IL-6 level in the blood of control mice, LPS-induced septic mice, and septic mice injected with sh-NC or sh-Peg13 lentivirus at 3, 7, and 14 days. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Figure 5 -. Up-regulation of Peg13 and HMGB1 in the blood of septic patients and negative correlation of Peg13 with HMGB1, CRP and ESR. (A) Peg13 expression in blood from healthy subjects and septic patients. (B) HMGB1 level in blood from healthy subjects and septic patients. (C) Peg13 expression is different between survivors and non-survivors. (D-F) Pearson correlation of Peg13 expression with HMGB1, CRP and ESR among septic patients. (G) Pearson correlation of lncRNA Peg13 expression with clinical SOFA score in patients with sepsis. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001;****p<0.0001.