Proteomic Analysis Of The Effect On Hepatic Stellate Cells With Herbal Compound 861

Hepatic fibrosis is a common response to chronic liver injury of variable origin (viral, metabolic, and toxic). It is the consequence of an imbalance between synthesis, deposition and degradation of extracellular matrix components Herbal compound 861(Cpd861) is an aqueous extract of 10 defined herbs with Salvia miltiorrhiza, Astragalus membranaceous, and Spatholobus suberectus as its chief components. Cpd861 has been shown experimentally and clinically to be effective in suppressing fibrogenesis, enhancing collagen degradation, and inhibiting tissue inhibitor of metalloproteinase (TIMP) expression. It is an effective compound against hepatic fibrosis and cirrhosis.To investigate the antifibrotic mechanism of Cpd861, we utilized the proteomics technology to the study of hepatic stellate cells treated with Cpd861.The rat hepatic stellate cell (HSC) was exposed to Cpd861 at a final concentration of 1mg/ml with control experiment. After incubation with Cpd861 for 48 hours, HSC was harvested and the total proteins were extracted. Two dimensional polyacrylamide gel electrophoresis is used to separate protein spots, and the good 2-DE pattern including high resolution and reproducibility was obtained. The stained gel images of Cpd861 group and normal control were analyzed with PDQUEST software.800-900 protein spots were detected. Average, of 910 and 890 protein spots were detected in 861 and control groups, respectively. The results showed that some protein spots displayed quantitative changes in expression. These proteins were digested with trypsin, measured with matrix-assisted laser desportion/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and searched in the database. 39 proteins that are up- or down- regulated in the 861 group and 29 proteins that are not distinctly differentially expressed are identified. We find some differential proteins,which correlate with liver fibrosis,collagen, regulatory proteins,and signal transduction. This investigation of HSCs treated with Cpd861 demonstrates that Cpd861 is efficient in suppressing the proliferation of HSCs, and accelerating the death of HSCs. Cpd861 may act against liver flbrosis in two ways of suppressing the signal transduction pathways involved with cellular collagen synthesis, and direct suppression synthesis of collagen synthesis.