Down-regulation of transforming growth factor beta 1/activin receptor-like kinase 1 pathway gene expression by herbal compound 861 is related to deactivation of LX-2 cells.
OBJECTIVE
To investigate the effect of herbal compound 861 (Cpd861) on the transforming growth factor-beta1 (TGF beta 1)/activin receptor-like kinase 1 (ALK1, type I receptor) signaling-pathway-related gene expression in the LX-2 cell line, and the inhibitory mechanism of Cpd861 on the activation of LX-2 cells.


METHODS
LX-2 cells were treated with TGF beta 1 (5 ng/mL) Cpd861 (0.1 mg/mL), TGF beta 1 (5 ng/mL) plus Cpd861 (5 ng/mL) for 24 h to investigate the effect of Cpd861 on the TGF beta 1/ALK1 pathway. Real-time PCR was performed to examine the expression of alpha-SMA (alpha-smooth muscle actin), ALK1, Id1 (inhibitor of differentiation 1). Western blotting was carried out to measure the levels of alpha-SMA and phosphorylated Smad1, and immunocytochemical analysis for the expression of alpha-SMA.


RESULTS
In LX-2 cells, TGF beta 1/ALK1-pathway-related gene expression could be stimulated by TGF beta 1, which led to excessive activation of the cells. Cpd861 decreased the activation of LX-2 cells by reducing the expression of alpha-SMA mRNA and protein expression. This effect was related to inhibition of the above TGF beta 1/ALK1-pathway-related expression of genes such as Id1 and ALK1, and phosphorylation of Smad1 in LX-2 cells, even with TGF beta 1 co-treatment for 24 h.


CONCLUSIONS
Cpd861 can restrain the activation of LX-2 cells by inhibiting the TGF beta 1/ALK1/Smad1 pathway.
Li
Li
L
Liver Research Center, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China.
Zhao
Xin-Yan
XY
Wang
Bao-En
BE
D000199:Actins / Q000378:metabolism; D030301:Activin Receptors, Type II / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D015536:Down-Regulation / Q000187:drug effects*; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D051798:Inhibitor of Differentiation Protein 1 / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D051898:Smad1 Protein / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
D000199:Actins; D004365:Drugs, Chinese Herbal; C497453:ID1 protein, human; D051798:Inhibitor of Differentiation Protein 1; C497399:SMAD1 protein, human; D051898:Smad1 Protein; D053773:Transforming Growth Factor beta1; C476499:herbal compound 861; C425364:ACVRL1 protein, human; D030301:Activin Receptors, Type II