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Pimpinellin ameliorates macrophage inflammation by promoting RNF146-mediated PARP1 ubiquitination
Liuye Yang1,2, Meng Du1,2,3,4, Kaiyuan Liu1,2
1Clinic Center of Human Gene Research, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Abstract:
Macrophage inflammation plays a central role during the development and progression of sepsis, while the regulation of macrophages by parthanatos has been recently identified as a novel strategy for anti-inflammatory therapies. This study was designed to investigate the therapeutic potential and mechanism of pimpinellin against LPS-induced sepsis. PARP1 and PAR activation were detected by western blot or immunohistochemistry. Cell death was assessed by flow cytometry and western blot. Cell metabolism was measured with a Seahorse XFe24 extracellular flux analyzer. C57, PARP1 knockout, and PARP1 conditional knock-in mice were used in a model of sepsis caused by LPS to assess the effect of pimpinellin. Here, we found that pimpinellin can specifically inhibit LPS-induced macrophage PARP1 and PAR activation. In vitro studies showed that pimpinellin could inhibit the expression of inflammatory cytokines and signal pathway activation in macrophages by inhibiting overexpression of PARP1. In addition, pimpinellin increased the survival rate of LPS-treated mice, thereby preventing LPS-induced sepsis. Further research confirmed that LPS-induced sepsis in PARP1 overexpressing mice was attenuated by pimpinellin, and PARP1 knockdown abolished the protective effect of pimpinellin against LPS-induced sepsis. Further study found that pimpinellin can promote ubiquitin-mediated degradation of PARP1 through RNF146. This is the first study to demonstrate that pimpinellin inhibits excessive inflammatory responses by promoting the ubiquitin-mediated degradation of PARP1.
Insights
Pimpinellin, a novel compound, combats sepsis by inhibiting macrophage inflammation. It targets Poly (ADP-ribose) polymerase 1 (PARP1) activation and promotes its degradation, offering a new therapeutic strategy for sepsis.
Area of Science:
- Immunology
- Pharmacology
- Molecular Biology
Background:
- Macrophage inflammation is critical in sepsis development and progression.
- Parthanatos regulation of macrophages presents a novel anti-inflammatory therapy target.
Purpose of the Study:
- To investigate the therapeutic potential and mechanism of pimpinellin in lipopolysaccharide (LPS)-induced sepsis.
- To explore pimpinellin's effect on macrophage activation and inflammatory pathways.
Main Methods:
- Western blot and immunohistochemistry to detect Poly (ADP-ribose) polymerase 1 (PARP1) and PAR activation.
- Flow cytometry and western blot for cell death assessment.
- Seahorse XFe24 extracellular flux analyzer for cell metabolism.
- In vivo studies using C57, PARP1 knockout, and PARP1 conditional knock-in mice models of LPS-induced sepsis.
Main Results:
- Pimpinellin specifically inhibited LPS-induced macrophage PARP1 and PAR activation.
- Pimpinellin reduced inflammatory cytokine expression and signal pathway activation in macrophages by inhibiting PARP1.
- Pimpinellin increased survival rates in LPS-treated mice and attenuated sepsis in PARP1 overexpressing mice.
- PARP1 knockdown abolished the protective effects of pimpinellin.
Conclusions:
- Pimpinellin demonstrates therapeutic potential against LPS-induced sepsis.
- Pimpinellin inhibits excessive inflammatory responses by promoting the ubiquitin-mediated degradation of PARP1 via RNF146.
- This study reveals a novel mechanism for pimpinellin's anti-inflammatory action in sepsis.
