α2-Macroglobulin reduces paracrine- and autocrine-stimulated matrix synthesis of cultured rat hepatic stellate cells

BACKGROUND: Transforming growth factor beta1 (TGF-beta1) is considered to represent a major fibrogenic mediator in the liver. The aim of the present study was to investigate whether alpha2-macroglobulin (alpha2M) might reduce paracrine- and autocrine-stimulated matrix synthesis of cultured rat hepatic stellate cells (HSCs) by scavenging TGF-beta. METHODS AND RESULTS: Using native agarose electrophoresis, we demonstrated that alpha2M binds [125I]-TGF-beta1 within minutes. Preincubation of transiently acidified supernatants of cultured Kupffer cells, secondary cultured (activated) HSC and platelet lysate with, respectively, 500 and 2000 microg mL-1 alpha2M significantly reduced the concentration of active TGF-beta1 in these media. As a consequence of TGF-beta scavenging by alpha2M, paracrine-stimulated proteoglycan synthesis of primary cultured HSCs was reduced significantly. Furthermore, addition of 200 microg mL-1 alpha2M to passaged (activated) HSCs resulted in (a) a reduction in autocrine-stimulated extracellular matrix synthesis (proteoglycan -52%, fibronectin -55%) and (b) increased cell proliferation. A similar reduction in matrix synthesis was observed after the addition of 5 micromol L-1 TGF-beta1 antisense oligonucleotide to activated HSCs. CONCLUSION: We conclude that alpha2M reduces paracrine-and autocrine-stimulated extracellular matrix synthesis of cultured HSCs by scavenging TGF-beta. These mechanisms might restrict liver fibrogenesis.