Abstract
Current researches have confirmed that Smads, mediators of TGF-β signaling, are strictly controlled by domain-specific site phosphorylation in the process of hepatic disease. Usually, Smad3 phospho-isoform pSmad3L and pSmad3C are reversible and antagonistic; pSmad2L/C could act together with pSmad3L by stimulating PAI-1 expression and ECM synthesis to transmit fibrogenic signals. Our recent study found that pSmad3C mutation is supposed to perform a vigorous role on the early phase of liver injury and abates salvianolic acid B’s anti-hepatic fibrotic-carcinogenesis. However, whether pSmad3C mutation expedites pSmad2L/C-mediated signaling transduction during hepatic fibrogenesis remains vague. Presently, Smad3 gene C-terminal phosphorylation site mutation heterozygote (pSmad3C+/-) mice were constructed to probe if and how pSmad3C retards CCl4-induced hepatic fibrogenesis by inhibiting pSmad2L/C-mediated signaling transduction. Twelve 6-week-old pSmad3C+/- C57BL/6J mice were intraperitoneally injection with CCl4 for 6 weeks to induce liver fibrogenesis. Results showed that pSmad3C mutation aggravates the relative liver weight, biochemical parameters, collagenous fibers and fibrotic septa formation, contributes to fibrogenesis in HT-CCl4 mice. Furthermore, fibrotic-related proteins TGF-β1, pSmad2C, pSmad2L, and PAI-1 were also increased in CCl4-induced pSmad3C+/- mice. These results suggest that pSmad3C mutation exacerbates hepatic fibrogenesis which relates to intensifying pSmad2L/C-mediated signaling transduction.
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Acknowledgements
We thank Prof. Koichi Matsuzaki for providing us with rabbit polyclonal anti-pSmad2L (Ser250/255) antibody.
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This work was supported by the National Natural Science Foundation of China (No. 82074073, No. 81874354, and No. 81573652).
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Contributors JY and YFG were responsible for conceptualization, investigation, and writing the original draft. WJX was responsible for resources and investigation and revised the manuscript. LLL was responsible for methodology, visualization, software, and analyzation of the data. ZHS was responsible for resources and investigation. QW and YHH were responsible for review and editing the manuscript. CZ, CCL, and ZRF were responsible for writing, review, and editing the manuscript. YY was responsible for the supervision and funding acquisition. All authors read and approved the manuscript, and all data were generated in-house and that no paper mill was used.
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Yang, J., Gong, Y., Xu, W. et al. Smad3 gene C-terminal phosphorylation site mutation exacerbates CCl4-induced hepatic fibrogenesis by promoting pSmad2L/C-mediated signaling transduction . Naunyn-Schmiedeberg's Arch Pharmacol 394, 1779–1786 (2021). https://doi.org/10.1007/s00210-021-02114-1
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DOI: https://doi.org/10.1007/s00210-021-02114-1