Suppression of CCl₄-induced liver fibrosis by the 34-mer.

<p>Liver fibrosis was induced by intraperitoneal injection of CCl₄ twice a week for 3 weeks. The mice then received the 34-mer twice a week intraperitoneally and were continuously injected with CCl₄ for further 4 weeks. (<b>A</b>) Histopathological detection of collagen in the liver by Sirius red-staining. (Original magnification, ×200). Representative pictures from at least 3 different experiments with six mice in each subgroup are shown. (<b>B</b>) Estimation of hepatic fibrosis area by Sirius red staining. Data were assessed by analyzing 10 Sirius red-stained liver sections per animal with a computerized image-plus system. *<i>P</i><0.02 <i>versus</i> CCl₄-treated 3 weeks-group; ^**<i>P</i><0.001 <i>versus</i> control peptide+CCl₄-treated group. (<b>C</b>) Relative mRNA expression of PDGF isoforms and receptors in CCl₄-treated mouse livers. CCl₄ treatment was performed as described as above. The cycle threshold (<i>Ct</i>) value of the PCR product and a control mRNA (GAPDH) were used to calculate the relative quantities of mRNA between samples. Data represent means ± SD of 6 animals in each group. *<i>P</i><0.05 versus untreated group. (D) Specimens were immunohistologically stained with α-SMA to observe activated HSCs (red color) and counterstained with Hoechst 33258 to observe nuclei. Original magnification, ×400. Scale bar = 20 µM. Numbers of α-SMA-positive cells are expressed as α-SMA-positive cells/400× field. Results were evaluated from 6 sections per liver specimen, and 6 mice in each group. ^**<i>P</i><0.005 <i>versus</i> control peptide+CCl₄-treated group. (<b>E</b>) qPCR analysis. The mRNA expression of TGFβ1, CoL1A1 and α-SMA was significantly attenuated by the 34-mer. The cycle threshold (<i>Ct</i>) value of the PCR product and a control mRNA (GAPDH) were used to calculate relative quantities of mRNA between samples. Data represent means ± SD of 6 animals in each group. **<i>P</i><0.05 versus control peptide+CCl₄-treated group.