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Updated: Apr 5, 2026
Cecal Ligation and Puncture-induced Sepsis as a Model To Study Autophagy in Mice
Published on: February 9, 2014
Selective Inhibition of SIRT2 Improves Outcomes in a Lethal Septic Model
1Department of Emergency, The First Hospital of China Medical University, 155 North Nanjing Street, Shenyang, Liaoning 110001, China. chongweixiena@yahoo.com.
Background:
Seven isoforms of histone deacetylase Class III have been reported - Sirtuin (SIRT) 1-7. We recently demonstrated that EX-527, an inhibitor of SIRT1, reduces mortality in a mouse model of lethal-cecal-ligationand- puncture (CLP)-induced septic shock. Our present study was aimed at determining whether selective inhibition of SIRT2, with AGK2, would decrease animal death and attenuate the inflammatory response in a septic model.
Methods:
Experiment I: C57BL/6J mice were intraperitoneally given either AGK2 (82 mg/kg) in dimethyl sulfoxide (DMSO) or DMSO alone, and 2 h later subjected to CLP. Survival was monitored for 240 hours. Experiment II: mice treated the same way as Experiment I, were grouped into (i) DMSO vehicle, and (ii) AGK2, with sham mice (operating but without any treatment) serving as controls. Peritoneal fluid and peripheral blood were examined at 24 and 48 hours for cytokine production. Samples of blood at 48 h were also allocated to assess coagulability using Thrombelastography (TEG). Morphological changes of bone marrow were evaluated from long bones (femurs and tibias) with hematoxylin and eosin (H&E) staining. Bone marrow atrophy was quantified by a blinded pathologist. Experiment III: cytokines in supernatant of the cultured normal primary splenocytes were measured after the cells were stimulated by lipopolysaccharide and treated with or without AGK2 (10 µM) for 6 hours.
Results:
AGK2 significantly reduced mortality and decreased levels of cytokines in blood (TNF-α: 298.3±24.6 vs 26.8±2.8 pg/ml, p=0.0034; IL-6: 633.4±82.8 vs 232.6±133.0 pg/ml, p=0.0344) and peritoneal fluid (IL-6: 704.8±67.7 vs 391.4±98.5 pg/ml, p=0.033) compared to vehicle control. Also, AGK2 suppressed the TNF-α and IL-6 production in the cultured splenocytes (TNF-α: 68.1±6.4 vs 23.9±2.8 pg/ml, p=0.0009; IL-6: 73.1±4.2 vs 49.6±3.0 pg/ml; p=0.0051). The TEG data showed that the mice subjected to CLP displayed prolonged fibrin formation and fibrin cross-linkage time, slower clot formation, decreased platelet function, and clot rigidity. AGK2 treatment was associated with dramatic improvements in fibrin cross-linkage and clot formation times, without a significant impact on the clot initiation parameters or platelet function. Additionally, AGK2 significantly attenuated the bone marrow atrophy (58.3±6.5 vs 30.0±8.2%, p=0.0262).
Conclusion:
Selective inhibition of SIRT2 significantly improves survival, and attenuates sepsis-associated "cytokine storm", coagulopathy, and bone marrow atrophy in a mouse model of lethal septic shock.
Insights
Selective inhibition of Sirtuin 2 (SIRT2) using AGK2 significantly improved survival in a mouse model of septic shock. AGK2 treatment reduced mortality, the inflammatory "cytokine storm," coagulopathy, and bone marrow atrophy.
Area of Science:
- Biochemistry
- Immunology
- Pharmacology
Background:
- Sirtuins (SIRTs) are a class of histone deacetylases with seven isoforms (SIRT1-7).
- SIRT1 inhibition with EX-527 reduced mortality in a mouse model of septic shock.
- The role of SIRT2 in sepsis pathophysiology remained largely unexplored.
Purpose of the Study:
- To investigate the therapeutic potential of selective SIRT2 inhibition using AGK2 in a mouse model of lethal septic shock.
- To determine if AGK2 treatment could decrease mortality and attenuate the inflammatory response during sepsis.
Main Methods:
- C57BL/6J mice were administered AGK2 or vehicle control before undergoing cecal ligation and puncture (CLP) to induce septic shock.
- Survival rates, cytokine levels (TNF-α, IL-6) in blood and peritoneal fluid, and splenocyte cytokine production were assessed.
- Coagulopathy was evaluated using Thrombelastography (TEG), and bone marrow atrophy was quantified via histological analysis.
Main Results:
- AGK2 treatment significantly reduced mortality in CLP-induced septic shock.
- AGK2 administration decreased circulating levels of TNF-α and IL-6, and suppressed their production in cultured splenocytes.
- Sepsis-induced coagulopathy and bone marrow atrophy were significantly attenuated by AGK2 treatment.
Conclusions:
- Selective inhibition of SIRT2 with AGK2 demonstrates significant therapeutic benefits in a preclinical model of septic shock.
- AGK2 effectively mitigates key pathological features of sepsis, including the "cytokine storm," coagulopathy, and bone marrow atrophy.
- Targeting SIRT2 represents a promising strategy for managing lethal septic shock.
