Figure 1.
In vivo effects of treatment with [44AANA47]-CCL5 on acute liver cell injury.
(A) Mice treated with [44AANA47]-CCL5 have reduced serum transaminases 48 hours after CCl4 application compared to vehicle treated mice. (B) Reduced liver cell damage is also evidenced by reduced necrotic areas 48 h hours after injury (HE staining, ×100). (C) The ameliorated damage after 48 h in [44AANA47]-CCL5 treated mice is preceded by a significantly reduced infiltration of CD45+ immune cells. (D) Reduced inflammatory liver damage in [44AANA47]-CCL5 is also associated with significantly repressed pro-inflammatory mRNA expression (black bars: [44AANA47]-CCL5, open bars: vehicle). * P<0.05, ** P<0.01.
Figure 2.
In vivo effects of treatment with [44AANA47]-CCL5 on chronic liver injury.
(A) Treatment of mice with [44AANA47]-CCL5 in parallel to chronic CCl4 intoxication reduced scar formation in the liver compared to vehicle (representative Sirius red stainings, ×100). Ameliorated liver fibrosis in [44AANA47]-CCL5 treated mice is quantified by a reduced Sirius red positive area (B) and a decreased hepatic concentration of the collagen specific amino acid hydroxyproline (C). ALT levels were not significantly changed in [44AANA47]-CCL5 treated mice compared to vehicle treated mice when taken 72 h after the last CCl4 injection (D). ** P<0.01.
Figure 3.
Treatment with [44AANA47]-CCL5 is associated with reduced stellate cell activation.
(A) Mice treated with [44AANA47]-CCL5 concomitantly to CCl4 display repressed mRNA expressions of the pivotal stellate cell genes Col1a1 and Timp1 compared to vehicle treated animals, while the mRNA of Tgf-β showed only a trend to lower expression. (B) Reduced activation of stellate cells in [44AANA47]-CCL5 treated mice is further evidenced by decreased protein expression of the stellate cell activation marker αSMA by immunohistochemistry (upper panels) and western bot of total liver extracts (lower panel). * P<0.05.
Figure 4.
Treatment with [44AANA47]-CCL5 leads to altered immune cell infiltration during liver fibrogenesis.
The photomicrographs show representative liver sections of mice treated with [44AANA47]-CCL5 or vehicle after staining for CD45 (total leukocytes), CD3 (T-cells) and macrophages (F4/80). Magnification ×100.
Figure 5.
Treatment with [44AANA47]-CCL5 leads to qualitative changes in splenocytes.
(A) Splenocytes were isolated from [44AANA47]-CCL5 and vehicle treated mice and conditioned mediate of splenocyte cultures were assessed for stellate cells migration in a Boyden chamber. Conditioned medium of splenocytes isolated from [44AANA47]-CCL5 treated mice (24 and 48 h after CCl4 application) was significantly less effective in inducing stellate cell migration compared to splenocytes isolated from vehicle treated mice. (B) Conditioned medium from mice treated [44AANA47]-CCL5 was also less effective in inducing stellate cell proliferation as assessed by BrdU incorporation. * P<0.05, ** P<0.01.