Leukamenin F suppresses liver fibrogenesis by inhibiting both hepatic stellate cell proliferation and extracellular matrix production
Abstract
Aim: To investigate the inhibitory effect of the natural product Leukamenin F on liver fibrosis and explore its potential underlying mechanisms.
Methods: Carbon tetrachloride (CCl4)-treated mouse model in vivo and in hepatic stellate cells (HSC) in vitro were used. The effect on CCl4-induced liver fibrosis was studied using histochemical and biochemical analysis, while the inhibition on HSC was assessed using cell proliferation/apoptosis assay and collagen I production using real-time PCR. The inhibitory effects of Leukamenin F on Akt/mTOR/p70S6K and TGFβ/Smad pathways was studied using Western blot and cell image analysis.
Results: Leukamenin F (0.1–1 mg/kg, ip, q.d.×28) significantly reduced α-SMA and collagen specific Sirius red staining areas in CCl4 -treated mouse livers. This compound at 1–2 μmol/L dose-dependently inhibited α-SMA expression, cell proliferation and type I procollagen mRNA expression in activated HSC. Furthermore it inhibited the Akt/mTOR/p70S6K pathway and suppressed TGFβ -induced Smad2/Smad3 phosphorylation and nuclear translocation in HSC.
Conclusion: Our results demonstrated that Leukamenin F could attenuate CCl4-induced liver fibrogenesis in mice as an efficient inhibitor against both HSC proliferation and ECM production. This natural product provides a valuable structural hint for the development of anti-liver fibrosis reagents.
Keywords:
Methods: Carbon tetrachloride (CCl4)-treated mouse model in vivo and in hepatic stellate cells (HSC) in vitro were used. The effect on CCl4-induced liver fibrosis was studied using histochemical and biochemical analysis, while the inhibition on HSC was assessed using cell proliferation/apoptosis assay and collagen I production using real-time PCR. The inhibitory effects of Leukamenin F on Akt/mTOR/p70S6K and TGFβ/Smad pathways was studied using Western blot and cell image analysis.
Results: Leukamenin F (0.1–1 mg/kg, ip, q.d.×28) significantly reduced α-SMA and collagen specific Sirius red staining areas in CCl4 -treated mouse livers. This compound at 1–2 μmol/L dose-dependently inhibited α-SMA expression, cell proliferation and type I procollagen mRNA expression in activated HSC. Furthermore it inhibited the Akt/mTOR/p70S6K pathway and suppressed TGFβ -induced Smad2/Smad3 phosphorylation and nuclear translocation in HSC.
Conclusion: Our results demonstrated that Leukamenin F could attenuate CCl4-induced liver fibrogenesis in mice as an efficient inhibitor against both HSC proliferation and ECM production. This natural product provides a valuable structural hint for the development of anti-liver fibrosis reagents.