Dataset Information

Costunolide represses hepatic fibrosis through WW domain-containing protein 2-mediated Notch3 degradation.

ABSTRACT:

Background and purpose

This study investigates the antifibrotic activities and potential mechanisms of costunolide (COS), a natural sesquiterpene compound.

Experimental approach

Rats subjected to bile duct ligation and mice challenged with CCl₄ were used to study the antifibrotic effects of COS in vivo. Mouse primary hepatic stellate cells (pHSCs) and human HSC line LX-2 also served as an in vitro liver fibrosis models. The expression of fibrogenic genes and signaling proteins in the neurogenic locus notch homologue protein 3 (Notch3)-hairy/enhancer of split-1 (HES1) pathway was examined using western blot and/or real-time PCR. Notch3 degradation was analysed using immunofluorescence and coimmunoprecipitation.

Key results

In animals, COS administration attenuated hepatic histopathological injury and collagen accumulation and reduced the expression of fibrogenic genes. COS time- and dose-dependently suppressed the levels of fibrotic markers in LX-2 cells and mouse pHSCs. Mechanistic studies showed COS destabilized Notch3 and subsequently inhibited the Notch3-HES1 pathway, thus inhibiting HSC activation. Furthermore, COS blocked the WW domain-containing protein 2 (WWP2)/protein phosphatase 1G (PPM1G) interaction and enhanced the effect of WWP2 on Notch3 degradation.

Conclusions and implications

COS exerted potent antifibrotic effects in vitro and in vivo by disrupting the WWP2/PPM1G complex, promoting Notch3 degradation and inhibiting the Notch3/HES1 pathway. This indicates that COS may be a potential therapeutic candidate for the treatment of liver fibrosis.

SUBMITTER: Ge MX

PROVIDER: S-EPMC6989949 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Publications

Costunolide represses hepatic fibrosis through WW domain-containing protein 2-mediated Notch3 degradation.

Ge Mao-Xu MX Liu Hong-Tao HT Zhang Na N Niu Wei-Xiao WX Lu Zhen-Ning ZN Bao Yun-Yang YY Huang Rui R Yu Dong-Ke DK Shao Rong-Guang RG He Hong-Wei HW

British journal of pharmacology 20191223 2

<h4>Background and purpose</h4>This study investigates the antifibrotic activities and potential mechanisms of costunolide (COS), a natural sesquiterpene compound.<h4>Experimental approach</h4>Rats subjected to bile duct ligation and mice challenged with CCl<sub>4</sub> were used to study the antifibrotic effects of COS in vivo. Mouse primary hepatic stellate cells (pHSCs) and human HSC line LX-2 also served as an in vitro liver fibrosis models. The expression of fibrogenic genes and signaling p ...[more]

PMID: 31621893

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Project description:The NLR family pyrin domain-containing 3 (NLRP3) inflammasome plays an important role in liver fibrosis (LF) development. However, the mechanisms involved in NLRP3-induced fibrosis are unclear. Our aim was to test the hypothesis that the NLRP3 inflammasome in hepatic stellate cells (HSCs) can directly regulate their activation and contribute to LF. Primary HSCs isolated from wild-type (WT), Nlrp3-/- , or Nlrp3L351PneoR knock-in crossed to inducible (estrogen receptor Cre-CreT) mice were incubated with lipopolysaccharide (LPS) and adenosine triphosphate (ATP), or 4OH-tamoxifen, respectively. HSC-specific Nlrp3L351P knock-in mice were generated by crossing transgenic mice expressing lecithin retinol acyltransferase (Lrat)-driven Cre and maintained on standard rodent chow for 6 months. Mice were then sacrificed; liver tissue and serum were harvested. Nlrp3 inflammasome activation along with HSC phenotype and fibrosis were assessed by RT-PCR, western blotting, fluorescence-activated cell sorting (FACS), enzyme-linked immunosorbent assay, immunofluorescence (IF), and immunohistochemistry (IHC). Stimulated WT HSCs displayed increased levels of NLRP3 inflammasome-induced reactive oxygen species (ROS) production and cathepsin B activity, accompanied by an up-regulation of mRNA and protein levels of fibrotic makers, an effect abrogated in Nlrp3-/- HSCs. Nlrp3L351P CreT HSCs also showed elevated mRNA and protein expression of fibrotic markers 24 hours after inflammasome activation induced with 4-hydroxytamoxifen (4OHT). Protein and mRNA expression levels of fibrotic markers were also found to be increased in isolated HSCs and whole liver tissue from Nlrp3L351P Lrat Cre mice compared to WT. Liver sections from 24-week-old NlrpL351P Lrat Cre mice showed fibrotic changes with increased alpha smooth muscle actin (?SMA) and desmin-positive cells and collagen deposition, independent of inflammatory infiltrates; these changes were also observed after LPS challenge in 8-week-old NlrpL351P Lrat Cre mice. Conclusion: Our results highlight a direct role for the NLRP3 inflammasome in the activation of HSCs directly triggering LF.

Dataset Information

Costunolide represses hepatic fibrosis through WW domain-containing protein 2-mediated Notch3 degradation.

Background and purpose

Experimental approach

Key results

Conclusions and implications

Publications

Costunolide represses hepatic fibrosis through WW domain-containing protein 2-mediated Notch3 degradation.

Similar Datasets

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