Abstract
Suppression of hepatic stellate cell (HSC) growth and activation, and induction of
apoptosis, have been proposed as therapeutic strategies for the treatment and prevention
of liver fibrosis. Our previous study showed that the Chinese herb Ligusticum chuanxiong (LC) inhibits platelet-derived growth factor (PDGF-BB)-induced HSC proliferation.
The present study was designed to investigate the active principles and their action
mechanisms. With a bioactivity-directed fractionation approach, DNA synthesis (bromodeoxyuridine
(BrdU) incorporation), cell cycle related proteins and apoptosis markers were determined
to evaluate the inhibitory effects of active principles of LC. Two phthalides, Z,Z′-6,8′,7,3′-diligustilide
(1) and levistolide A (2), from LC significantly abrogated PDGF-BB-induced proliferation in both rat and human
HSC lines. These inhibitory effects of compounds 1 and 2 were associated with reduction of α-smooth muscle actin and collagen expressions.
The cell cycle promoting proteins, cyclins D1, D2, E, A and B1, were downregulated
while the inhibitory proteins p21 and 27 were up-regulated. JNK phosphorylation was
up-regulated by compounds 1 and 2. In HSC-T6, the two compounds induced apoptosis through the activation of caspases
9 and 3, increase in cytosolic cytochrome c release, and downregulation of Bcl-2 and Akt phosphorylation. Moreover, neither phthalides
caused direct cytotoxicity to either HSCs or rat primary hepatocytes under experimental
concentrations. These results indicate that two phthalides from LC inhibited PDGF-BB-activated
HSC proliferation possibly through cell cycle inhibition and apoptosis mechanisms.
They might be potential anti-fibrotic drugs for the treatment and prevention of hepatic
fibrosis.
Abbreviations
BrdU:bromodeoxyuridine
EtOAc:ethyl acetate
ECM:extracellular matrix
HSC:hepatic stellate cell
JNK:c-jun N-terminal kinase
LC:Ligusticum chuanxiong
NAC:N-acetylcysteine
PDGF:platelet-derived growth factor
α-SMA:α-smooth muscle actin
TNF:tumor necrosis factor
Key words
Hepatic stellate cells -
Ligusticum chuanxiong
- Umbelliferae - platelet-derived growth factor - phthalides - apoptosis
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Dr. Yi-Tsau Huang
Institute of Traditional Medicine
School of Medicine
National Yang-Ming University
No. 155, Li-Nong Street, Sect. 2
Taipei 112
Taiwan
Email: huangyt@ym.edu.tw
