Objective    To investigate the effects of tanshinol on the proliferation

apoptosis and NF-κB activation in rat hepatic stellate cells (HSCs) after IL-1β inducement

a

nd to elucidate the anti-fibrotic molecular mechanisms of tanshinol.     Methods    The rat HSCs was isolated with collagenase

in situ

liver recirculation perfusion and cultured

in vitro

. The cells were divided into 5 groups: normal control

IL-1β treatment group (10 ng/ml)

and tanshinol group 1

2 and 3. The later 3 groups were pretreated with tanshinol at the concentrations of 0.062 5

0.125 and 0.25 mmol/L respectively followed by 10 ng/ml IL-1β treatment 1 h later. MTT colorimetric assay was used to detect the proliferation of HSCs. AO/EB immunoflurorescence microscopy and combination Annexin-V-FITC/PI double-labelimmunofluorescence with flow cytometer were employed to examine the apoptosis of HSCs. Synthesis and secretion of collagen Ⅲ were detected by the quantitative immunocytochemical assay and ELISA respectively. The amounts of cytoplasm p-IκBα and NF-κB p65

and nuclear NF-κB p65 in HSCs were determined by Western blotting. Immunocytochemical staining and Western blotting were used to observe nuclear translocation of NF-κB p65.     Results    IL-1β increased the proliferation of HSCs (

P

<

0.01)

reduced the cell apoptosis (

P

<

0.05)

and facilitated the synthesis and secretion of collagen Ⅲ in HSCs (

P

<

0.05

P

<

0.01) in comparison with normal control. After treated with different concentration of tanshinol

HSC’s proliferation was prominently reduced (

P

<

0.01) and its apoptosis was increased (

P

<

0.01)

especially for early apoptosis. Tanshinol also degraded the synthesis and secretion of collagen Ⅲ in HSCs

especially in 0.25 mmol/L concentration group (

P

<

0.01). Western blotting showed that IL-1β elevated amounts of cytoplasm p-IκBa and nuclear NF-κB p65 (

P

<

0.01)

decreased amounts of cytoplasm p65 markedly (

P

<

0.05)

then facilitated nuclear translocation of p65. Immunocytochemical staining indicated that the number of nuclear NF-κB p65 positive HSCs were increased and the colo

r was deepened. IL-1β induced NF-κB activation in rat HSCs. Tanshinol reduced the amounts of cytoplasm p-IκBa and nuclear NF-κB p65 (

P

<

0.05

P

<

0.01)

and decreased nuclear translocation of p65 markedly. Immunocytochemical staining showed that the number of nuclear NF-κB p65 positive HSCs was decreased and the color grew pallid. Above results indicated that tanshinol inhibited IL-1β-induced NF-κB activation in rat HSCs.     Conclusion    Tanshinol inhibits the proliferation of HSCs

facilitates the cell apoptosis

and reduces the synthesis and secretion of collagen   Ⅲ  through inhibiting NF-κB activation

via

suppressing hepatic fibrosis.