N-(Methylamino) Isobutyric Acid Inhibits Proliferation of CFSC-2C Hepatic Stellate Cells

Authors

T. L. Freeman, Veterans Administration Medical Center, Alcohol Study Unit, Omaha, NE, USA
G.M. Thiele, Veterans Administration Medical Center, Alcohol Study Unit, Omaha, NE, USAFollow
L.W. Klassen, Veterans Administration Medical Center, Alcohol Study Unit, Omaha, NE, USA
B.T. Klassen, Veterans Administration Medical Center, Alcohol Study Unit, Omaha, NE, USA
M.E. Mailliard, Veterans Administration Medical Center, Alcohol Study Unit, Omaha, NE, USA

Date of this Version

2004

Comments

Published in Biochemical Pharmacology 68 (2004) 223–230.

Abstract

Activation of hepatic stellate cells (HSCs) involves the induction of ECM protein synthesis and rapid cell proliferation. Thus, agents that interfere with either process could potentially mitigate the development of liver disease by reducing the synthesis of proteins associated with fibrosis or by reducing the number of activated HSC. Previously, we described that the non-metabolizable amino acid analog N-(methylamino)isobutyric acid (MeAIB) reduced hepatic collagen content of rats in a model of CCl₄-induced liver injury, and in vitro studies using CFSC-2G cells indicated that MeAIB directly reduced collagen synthesis. However, the MeAIB-mediated reduction of hepatic collagen, in vivo, following liver injury was associated with a decrease in hepatic α-smooth muscle actin (α-SMA) which suggested that MeAIB also inhibited the activation of HSCs. Because HSC activation is inseparable from proliferation, the purpose of this study was to examine the effect of MeAIB treatment on the proliferation of HSCs in an in vitro model utilizing CFSC-2G cell cultures. In these studies, MeAIB effectively inhibited the proliferation of CFSC-2G cells by interfering with the progression of the cells through the G₁-phase of the cell cycle which delayed entry into S-phase. MeAIB prevented the phosphorylation of p70S6 kinase (p70S6K) at Thr389 and reduced the phosphorylation at Thr421/Ser424. Because p70S6K is required for G₁-cell cycle progression and is known to be regulated by nutrient availability, this correlates well with MeAIB interfering with the proliferation of CFSC-2G HSCs. In addition, the rate of protein synthesis was reduced by MeAIB treatment following mitogenic stimulation, which agrees with a p70S6K-mediated reduction in translation. These data are consistent with MeAIB inhibiting the proliferation of CFSC-2G cells by altering the mitogen activated pathway(s) leading to phosphorylation of p70S6K by a yet to be described mechanism.