Objective:

2

To explore the effect and mechanism of Dahuang Zhechong Pill (DHZCP) on liver fibrosis.

Methods:

2

Liver fibrosis cell model was induced by transforming growth factor-β (TGF-β) in hepatic stellate cells (HSC-T6). DHZCP medicated serum (DMS) was prepared in rats. HSC-T6 cells were divided into the control (15% normal blank serum culture)

TGF-β (15% normal blank serum + 5 ng/mL TGF-β)

DHZCP (15% DMS + 5 ng/mL TGF-β)

DHZCP+PDTC [15% DMS + 4 mmol/L ammonium pyrrolidine dithiocarbamate (PDTC)+ 5 ng/mL TGF-β]

and PDTC groups (4 mmol/L PDTC + 5 ng/mL TGF-β). Cell activity was detected by cell counting kit 8 and levels of tumor necrosis factor-α (TNF-α)

interleukin (IL)-1β

IL-6

aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the cell supernatant were determined by enzyme-linked immunosorbnent assay. Western blot was used to measure the expressions of p38 mitogen-activated protein kinase/nuclear factor kappa B/transforming growth factor-β1 (p38 MAPK/NF-κB/TGF-β1) pathway related proteins

and the localization and expressions of these proteins were observed by immunofluorescence staining.

Results:

2

DHZCP improved the viability of cells damaged by TGF-β and reduced inflammatory cytokines and ALT and AST levels in the supernatant of H

SC-T6 cells induced with TGF-β (

P

<

0.05 or

P

<

0.01). Compared with the TGF-β group

NF-κB p65 levels in the DHZCP group were decreased (

P

<

0.05). p38 MAPK and NF-κB p65 levels in the DHZCP+PDTC were also reduced (

P

<

0.01). Compared with the TGF-β group

the protein expression of Smad2 showed a downward trend in the DHZCP

DHZCP+PDTC

and PDTC groups (all

P

<

0.01)

and the decreasing trend of Samd3 was statistically significant only in DHZCP+PDTC group (

P

<

0.01)

whereas Smad7 was increased (

P

<

0.05 or

P

<

0.01).

Conclusion:

2

DHZCP can inhibit the process of HSC-T6 cell fibrosis by down-regulating the expression of p38 MAPK/NF-κB/TGF-β1 pathway.