Dataset Information

Withagulatin A inhibits hepatic stellate cell viability and procollagen I production through Akt and Smad signaling pathways.

ABSTRACT:

Aim

To investigate the effects of the natural product Withagulatin A on hepatic stellate cell (HSC) viability and type I procollagen production. The potential mechanism underlying the pharmacological actions was also explored.

Methods

The effect of Withagulatin A on cell viability was evaluated in HSC and LX-2 cells using a sulforhodamine B (SRB) assay. Cell cycle distribution was analyzed using flow cytometry. Type I procollagen gene expression was determined using real-time PCR. Regulation of signaling molecules by Withagulatin A was detected using Western blotting.

Results

Primary rat HSCs and the human hepatic stellate cell line LX-2 treated with Withagulatin A (0.625-20 micromol/L) underwent a dose-dependent decrease in cell viability, which was associated with S phase arrest and the induction of cell apoptosis. In addition, the natural product decreased phosphorylation of the Akt/mTOR/p70S6K pathway that controls cell proliferation and survival. Furthermore, Withagulatin A (1, 2 mumol/L) inhibited transforming growth factor-beta (TGF-beta) stimulated type I procollagen gene expression, which was attributable to the suppression of TGF-beta stimulated Smad2 and Smad3 phosphorylation.

Conclusion

Our results demonstrated that Withagulatin A potently inhibited HSC viability and type I procollagen production, thereby implying that this natural product has potential use in the development of anti-fibrogenic reagents for the treatment of hepatic fibrosis.

SUBMITTER: Liu Q

PROVIDER: S-EPMC4007817 | biostudies-literature | 2010 Aug

REPOSITORIES: biostudies-literature

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Publications

Withagulatin A inhibits hepatic stellate cell viability and procollagen I production through Akt and Smad signaling pathways.

Liu Qiong Q Chen Jing J Wang Xu X Yu Liang L Hu Li-hong LH Shen Xu X

Acta pharmacologica Sinica 20100719 8

<h4>Aim</h4>To investigate the effects of the natural product Withagulatin A on hepatic stellate cell (HSC) viability and type I procollagen production. The potential mechanism underlying the pharmacological actions was also explored.<h4>Methods</h4>The effect of Withagulatin A on cell viability was evaluated in HSC and LX-2 cells using a sulforhodamine B (SRB) assay. Cell cycle distribution was analyzed using flow cytometry. Type I procollagen gene expression was determined using real-time PCR. ...[more]

PMID: 20644552

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Project description:BackgroundIn mice, liver fibrosis is the most serious pathologic change during Schistosoma japonicum (S. japonicum) infection. Schistosomiasis is mainly characterized by schistosome egg-induced granulomatous fibrosis. Hepatic stellate cells (HSCs) are mainly responsible for the net accumulation of collagens and fibrosis formation in the liver. Activated HSCs regulated by transforming growth factor-β1 (TGF-β1)/Smad signaling have emerged as the critical regulatory pathway in hepatitis virus or carbon tetrachloride-induced liver fibrosis. However, the detailed mechanism of HSC activation in schistosome-induced liver fibrosis is poorly understood.MethodsSchistosoma japonicum-induced murine models and a control group were generated by abdominal infection with 15 (± 1) cercariae. The purity of cultured primary HSCs was evaluated by immunocytochemistry. The histopathological changes in the livers of infected mice were estimated by hematoxylin-eosin and Masson staining. Dynamic expression of pro-fibrotic molecules and microRNAs was detected by real-time quantitative PCR (RT-qPCR). Mainly members involved in the TGF-β1/Smad signaling pathway were examined via RT-qPCR and Western blot.ResultsThe egg-induced granulomatous inflammation formed at 4 weeks post-infection (wpi) and developed progressively. Alpha-smooth muscle actin (α-SMA), collagen I, collagen III, TGF-β1, Smad2, Smad3, and Smad4 showed a significant increase in mitochondrial RNA (mRNA) and protein expression compared with the control group at 7 and 9 weeks post-infection (wpi), while an opposite effect on Smad7 was observed. In addition, the mRNA expression of microRNA-21 (miRNA-21) was significantly increased at 7 wpi, and the mRNA expression of miRNA-454 was decreased starting from 4 wpi.ConclusionOur present findings revealed that HSCs regulated by the TGF-β1/Smad signaling pathway play an important role in liver fibrosis in S. japonicum-infected mice, which may provide proof of concept for liver fibrosis in schistosomiasis.

Dataset Information

Withagulatin A inhibits hepatic stellate cell viability and procollagen I production through Akt and Smad signaling pathways.

Aim

Methods

Results

Conclusion

Publications

Withagulatin A inhibits hepatic stellate cell viability and procollagen I production through Akt and Smad signaling pathways.

Similar Datasets

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