Early genes induced in hepatic stellate cells during wound healing.

Activation of mesenchymal cells is a central event in the wound healing response of most tissues. In liver, the mesenchymal element responsible for organ fibrosis is the hepatic stellate cell (HSC) (formerly known as lipocyte or Ito cell). The phenotypic cascade of stellate cell activation in liver fibrosis has been well documented and involves both marked morphologic changes and upregulation of several functional components including extracellular matrix, cytokine receptors, contractile filaments and metalloproteinases. However, the genetic regulation of stellate cell activation is poorly understood. In an attempt to clone genes that are involved in the regulation of HSC activation we have combined cDNA library amplification by PCR with subtraction hybridization/differential screening, and have successfully identified genes induced in vivo during early stellate cell activation in a rat model of liver fibrosis. The subtracted cDNA library comprised less than 100 unique sequences. Of these, 13 clones with sizes ranging from 322 to 745 were sequenced and characterized. Gene induction in HSCs was monitored by RNAse protection assay during early liver injury induced by the hepatotoxin CCl4. The sequenced cDNAs corresponding to the known genes included type II transforming growth factor beta receptor, glutathione peroxidase I, transferrin and several clones encoding cellular retrotransposons, whose expression was not previously identified in non-parenchymal liver cells. In addition, one partial cDNA predicted a zinc-finger motif, suggesting a possible role of a novel transcriptional regulator. Our approach represents a valuable strategy for clarifying in vivo regulatory mechanisms of mesenchymal cell activation in wound healing.