Effects of p-CREB-1 on transforming growth factor-β3 auto-regulation in hepatic stellate cells.

Previous studies have demonstrated that transforming growth factor-β3 (TGF-β3) protected liver against fibrosis in vivo and vitro, but its regulation is poorly understood. In addition, the cAMP-responsive element (CRE) in TGF-β3 promoter is recognized as an important regulatory site for TGF-β3 auto-regulation. Thus, we hypothesize that transcription factor CRE-binding protein-1 (CREB-1) regulates the auto-induction of TGF-β3 in hepatic stellate cells (HSCs). We used exogenous TGF-β3 to activate the signal pathway of TGF-β3 auto-regulation in HSCs, results indicated that exogenous TGF-β3 could up-regulate the protein and mRNA expressions of TGF-β3, and provoke the phosphorylation of CREB-1 on Ser-133, besides, it could induce the DNA binding activity of p-CREB-1 and activate TGF-β3 promoter as well. Additionally, we used pGenesil-1.1-shRNA-CREB-1 and pRSV-CREB-1 expression vector to silence and up-regulate CREB-1 gene expression respectively, and the results indicated that inhibition of CREB-1 suppressed exogenous TGF-β3 stimulation of TGF-β3 mRNA and protein expressions in HSCs, whereas up-regulation of CREB-1 induced this stimulation. Our results indicate that exogenous TGF-β3 up-regulates the activity of TGF-β3 promoter by activating CREB-1, then induces the mRNA and protein expressions of TGF-β3. Especially, p-CREB-1 is a critical transcription factor in mediating TGF-β3 auto-induction.