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Updated: Aug 1, 2025
Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies
Published on: April 11, 2012
Bioactive compound C498-0670 alleviates LPS-induced sepsis via JAK/STAT and NFκB signaling pathways
Jing Xu1,2, Xinxin Zhang2, Mingming Zhou2
1Department of Gastroenterology, The Affiliated Hospital of Qingdao University, Qingdao, China.
Abstract:
The JAK/STAT and NFκB signaling pathways are two major inflammatory signaling pathways that are usually activated simultaneously in the body's inflammatory response to bacterial or viral infections. Hyperactivation of these two prominent signaling pathways is associated with various immune-related diseases and mortality, pointing to an urgent need for drug development targeting JAK/STAT and/or NFκB signaling. In this study, we screened 18,840 compounds using our well-established dual STAT-NFκB driven luciferase reporter based high-throughput screening system and identified a bioactive compound C498-0670, which inhibits both JAK/STAT and NFκB signaling. C498-0670 inhibits the activation of STATs and p-IKKα/β in both the immortalized cell lines and primary peritoneal macrophages, while suppressing the expression of LPS-induced inflammatory mediators in vitro. In addition, the overall anti-inflammatory effects of C498-0670 were investigated using transcriptome sequencing and bioinformatics approaches. C498-0670 was predicted to alleviate sepsis/septic shock by disease/function analysis using IPA software, which was further verified in the LPS-induced mouse sepsis model in vivo. C498 reduced LPS-induced liver and kidney damage, myeloid cell infiltration, and pro-inflammatory cytokine and chemokine production in vivo. Furthermore, the SPR-HPLC-MS-based target fishing approach was used to identify the putative drug targets, and the high affinities of JAK2 (JAK/STAT signaling), NFKBIA (NFκB signaling), and IL-1β, NLRP1b (inflammasome signaling) for C498-0670 were verified by molecular docking approach. These results suggest that C498-0670 can be used as a dual-target inhibitor of JAK/STAT and NFκB signaling pathways for the treatment of various inflammatory diseases, especially septic shock.
Insights
Researchers identified C498-0670, a novel compound inhibiting both Janus kinase/signal transducer and activator of transcription (JAK/STAT) and nuclear factor kappa B (NFκB) pathways. This dual-action inhibitor shows promise for treating inflammatory diseases like septic shock.
Area of Science:
- Immunology
- Pharmacology
- Molecular Biology
Background:
- The Janus kinase/signal transducer and activator of transcription (JAK/STAT) and nuclear factor kappa B (NFκB) signaling pathways are critical in inflammatory responses.
- Hyperactivation of these pathways is linked to immune-related diseases and increased mortality.
- Targeting JAK/STAT and/or NFκB signaling is crucial for developing new anti-inflammatory therapeutics.
Purpose of the Study:
- To identify novel compounds that inhibit both JAK/STAT and NFκB signaling pathways.
- To evaluate the therapeutic potential of identified compounds in inflammatory conditions, particularly septic shock.
Main Methods:
- High-throughput screening of 18,840 compounds using a dual STAT-NFκB luciferase reporter system.
- In vitro validation of compound activity in cell lines and primary macrophages.
- Transcriptome sequencing and bioinformatics analysis for anti-inflammatory effects.
- In vivo studies using a lipopolysaccharide (LPS)-induced mouse sepsis model.
- Target fishing (SPR-HPLC-MS) and molecular docking to identify drug targets.
Main Results:
- A bioactive compound, C498-0670, was identified that inhibits both JAK/STAT and NFκB signaling.
- C498-0670 suppressed STATs activation, p-IKKα/β, and inflammatory mediator expression in vitro.
- In vivo, C498-0670 alleviated LPS-induced liver and kidney damage, reduced myeloid cell infiltration, and decreased pro-inflammatory cytokine/chemokine production.
- JAK2, NFKBIA, IL-1β, and NLRP1b were identified as potential targets with high binding affinity for C498-0670.
Conclusions:
- C498-0670 acts as a dual-target inhibitor of JAK/STAT and NFκB signaling pathways.
- The compound demonstrates significant anti-inflammatory effects in vitro and in vivo, particularly in a septic shock model.
- C498-0670 holds potential for treating inflammatory diseases, including septic shock.
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