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| The mechanism of action of total glycosides of paeony in attenuating liver fibrosis based on TGF-β/Smad and Fas/FADD signaling pathways |
| Hits 376 Download times 128 Received:May 27, 2025 |
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| DOI
10.11656/j.issn.1672-1519.2025.10.12 |
| Key Words
Paeoniae Radix Alba;total glycosides of paeony;hepatic fibrosis;apoptosis;hepatic stellate cells |
| Author Name | Affiliation | E-mail | | FAN Xiaoxu | Beijing University of Chinese Medicine, Beijing 100029, China | | | LIU Shuangqiao | North Sichuan Medical College, Nanchong 637100, China | | | HUA Ji'an | Beijing University of Chinese Medicine, Beijing 100029, China | | | XIE Zhijiu | Xinjiang Medical University, Urumqi 830011, China | | | WANG Zhen | Beijing University of Chinese Medicine, Beijing 100029, China | | | SHEN Yiwei | Beijing University of Chinese Medicine, Beijing 100029, China | | | JIN Zhenhui | Beijing University of Chinese Medicine, Beijing 100029, China | | | WANG Jingxia | Beijing University of Chinese Medicine, Beijing 100029, China | wjx20131210@163.com |
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| Abstract
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| Objective To investigate the mechanism of action of total glucosides of paeony(TGP) on TGF-β1-induced fibrosis in activated HSC-LX2 cells based on transforming growth factor-β(TGF-β)/Smad and Fas/Fas-associated with death domain protein(FADD) signaling pathways.Methods Hepatic fibrosis model was established using 10 ng/mL TGF-β1 modeling drug working solution to induce HSC-LX2 cell activation. Cells were intervened with different concentrations of TGP(125, 250, 500 μg/mL) for 48 h. The cell survival rate of each group was detected by cell viability assay kit(CCK-8), the apoptosis rate of each group was detected by flow cytometry, and the inflammatory factors COX2, IL-1β, and TNF-α content were detected by enzyme-linked immunosorbent assay(ELISA), and Collagen I(COL-Ⅰ), Collagen Ⅲ(COL-Ⅲ), and α-smooth muscle actin(α-SMA) protein expressions were detected by protein immunoblotting(Western blot). These are to determine whether the model is valid and TGP efficacy. Secondly, protein expression of tissue inhibitor of metal protease 1(TIMP1) and matrix metallopeptidase 1(MMP1) was detected to determine the degree of extracellular matrix deposition even further. Subsequently, TGF-β1, Smad2, Smad3, Fas, FADD, Caspase-3, and Caspase-8 mRNA and protein expression were detected to reveal the possible mechanisms by which TGP regulates hepatocyte fibrosis through TGF-β/Smad and Fas/FADD signaling pathways.Results After 48 h of TGP intervention, compared with the blank group, HSC-LX2 cells in the model group showed activation and proliferation, elevated COL-Ⅰ, COL-Ⅲ, α-SMA, and TIMP1 protein expression, and decreased MMP1 protein expression(P < 0.05), indicating that the hepatocyte fibrosis model was established. Compared with the model group, 125, 250, and 500 μg/mL TGP significantly inhibited activated HSC-LX2 cell activity(P < 0.05), enhanced apoptosis rate(P < 0.001), and decreased COX2, IL-1β, and TNF-α content(P < 0.01 or P < 0.001); 250 μg/mL and 500 μg/mL TGP decreased COL-Ⅰ and TIMP1 protein expression(P < 0.05) and increased MMP1 protein expression level(P < 0.05); 500 μg/mL TGP decreased COL-Ⅲ and α-SMA protein expression(P < 0.05);various concentrations of TGP inhibited TGF-β1, Smad2, and Smad3 mRNA and protein expression in the TGFβ/Smad signaling pathway and stimulated Fas, FADD, Caspase-3, and Caspase-8 mRNA and protein expression in the Fas/FADD signaling pathway(P < 0.05) to varying degrees.Conclusion The mechanism of TGP-induced fibrotic action in activated HSC-LX2 cells may be through the inhibition of the TGF-β/Smad signaling pathway to alleviate collagen fiber accumulation and extracellular matrix deposition in the process of hepatocyte fibrosis, whereas the mechanism of prompting apoptosis in activated HSC-LX2 cells may be through the up-regulation of the Fas/FADD signaling pathway. |
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