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Transforming growth factor beta gene expression is transiently enhanced at a critical stage during liver regeneration
J Armendariz-Borunda1, H Katai, C M Jones
1Veterans Affairs Medical Center, Nashville, Tennessee.
Background:
Transforming growth factor beta 1 (TGF beta 1) gene expression is increased in CCl4-injured rat livers. The biologic link of this increase and liver regeneration has not been established.
Experimental Design:
To explore the identity of the TGF beta 1-producing cells in the CCl4 regenerating liver, we hybridized untreated and CCl4-treated liver sections of TGF beta 1-specific riboprobes and immunolocalized TGF beta 1 protein, simultaneously. To assess the dynamics of cellular proliferation during hepatic regeneration, chronologic changes in the cellular DNA synthesis were also monitored by [3H]thymidine incorporation and autoradiography. In situ hybridization analyses were further extended by subfractionation of nonparenchymal cells into Kupffer cells/macrophages, endothelial and Ito cells, and determining TGF beta 1 mRNA levels in different cell types. Finally, we experimentally tested if the temporal relationship between the transient elevation of expression of TGF beta 1 and hepatic cell proliferation were casually related.
Results:
We observed that the rate of DNA synthesis was the highest around 36 hours post-treatment and preceded the time of enhanced accumulation of TGF beta 1 transcripts and protein, both of which peaked at approximately 48 hours and declined thereafter. Transient upregulation of TGF beta 1 gene expression was seen in the inflammatory cell infiltrates around the central vein and at less extent, in portal tracts, and in perisinusoidal cells near the zone of necrosis. Like TGF beta 1 transcripts, TGF beta 1 protein was also predominantly co-localized in and around the pericentral and periportal cells. Kupffer cells, that accumulate abundantly in the liver 48 hours after CCl4 administration, were the primary producers of TGF beta 1. The injection of neutralizing anti-TGF beta 1 antibodies into animals prevented both the decline in [3H]thymidine incorporation and cell division in the waning phases of hepatic regeneration at 72 hours.
Conclusions:
Based on our observations that (i) TGF beta 1 gene expression is triggered transiently during a crucial phase of liver regeneration, (ii) the exogenously added TGF beta 1 inhibits hepatic DNA synthesis and that (iii) the administration of TGF beta 1 antibodies extends the proliferative response of the regenerating liver, we conclude that TGF beta 1 plays a pivotal role in down regulating liver regeneration.