Establishment of assessment system for anti-fibrotic activity using an ex vivo hepatic fibrosis model

Abstract

Currently, there is no effective treatment for liver fibrosis. In vitro studies on the activation of hepatic stellate cells (HSCs), which is responsible for liver fibrosis, have been used as a drug screening system for it. However, even if some drug shows an inhibitory effect on HSC activation, its anti-fibrotic effect is required to be confirmed in liver fibrosis animal models, which further requires a lot of time and cost. In the present study, we tried to establish an ex vivo model of liver fibrosis using precision-cut liver slices (PCLSs) to solve such problems. PCLSs of 250 µm thickness were prepared from male C57BL/6J mice using a vibratome and cultured in RPMI medium in a 5% CO 2 incubator. Although cellular ATP content was decreased on day 1 compared to day 0, it was then maintained until day 5, suggesting that the ex vivo model is viable for at least 5 days. Treatment with Et-OH (50, 100 mM), one of the liver injury stimuli, for 5 days increased mRNA expression of Acta2 and Col1a1, liver fibrosis markers, in PCLSs. DIF-1 (50, 100 µM), which has an anti-fibrotic effect, significantly suppressed the Et-OH-induced increases in the markers. These results suggest that the ex vivo model using PCLSs is useful as a drug screening system for the development of drugs for treatment of liver fibrosis.