Less Severe Sepsis in Cecal Ligation and Puncture Models with and without Lipopolysaccharide in Mice with Conditional Ezh2-Deleted Macrophages (LysM-Cre System)

Abstract

Despite a previous report on less inflammatory responses in mice with an absence of the enhancer of zeste homologue 2 (Ezh2), a histone lysine methyltransferase of epigenetic regulation, using a lipopolysaccharide (LPS) injection model, proteomic analysis and cecal ligation and puncture (CLP), a sepsis model that more resembles human conditions was devised. As such, analysis of cellular and secreted protein (proteome and secretome) after a single LPS activation and LPS tolerance in macrophages from Ezh2 null (Ezh2flox/flox; LysM-Crecre/−) mice (Ezh2 null) and the littermate control mice (Ezh2fl/fl; LysM-Cre−/−) (Ezh2 control) compared with the unstimulated cells from each group indicated fewer activities in Ezh2 null macrophages, especially by the volcano plot analysis. Indeed, supernatant IL-1β and expression of genes in pro-inflammatory M1 macrophage polarization (IL-1β and iNOS), TNF-α, and NF-κB (a transcription factor) were lower in Ezh2 null macrophages compared with the control. In LPS tolerance, downregulated NF-κB compared with the control was also demonstrated in Ezh2 null cells. In CLP sepsis mice, those with CLP alone and CLP at 2 days after twice receiving LPS injection, representing sepsis and sepsis after endotoxemia, respectively, symptoms were less severe in Ezh2 null mice, as indicated by survival analysis and other biomarkers. However, the Ezh2 inhibitor improved survival only in CLP, but not LPS with CLP. In conclusion, an absence of Ezh2 in macrophages resulted in less severe sepsis, and the use of an Ezh2 inhibitor might be beneficial in sepsis.

Kazachstania pintolopesii in Blood and Intestinal Wall of Macrophage-Depleted Mice with Cecal Ligation and Puncture, the Control of Fungi by Macrophages during Sepsis

“…The control group was injected with liposome control. One week after injection, cecal ligation puncture (CLP) or sham surgery were performed according to an established protocol [25][26][27]. Briefly, the cecum was ligated at 1 cm from the cecal tip, punctured twice with a 21-gauge needle, and gently squeezed to release a small amount of fecal material in the peritoneum.…”

“…Kidney injury scores were determined by tubular cell damage with the scores as previously published [31,32], including, 0; no damage, 1; 0-25% tubular damage, 2; 25-50% tubular damage, 3; 50-75% tubular damage, and 4; 75-100% tubular damage. Liver damage was determined based on inflammation and hepatocyte necrosis with the following scores, including 0 (no finding); 1 (mild); 2 (moderate); and 3 (severe) [25,[33][34][35]. Colon damage scores were evaluated using the Geboes Score (GS) based on the erosions or ulcerations, mononuclear cell infiltration, and structural alterations with scores ranging from 0-5 [36].…”