Hepatocyte cytoskeleton during ischemia and reperfusion - influence of ANP-mediated p38 MAPK activation

Figure 6 Influence of ischemia and ANP pretreatment on Hsp27 distribution. After perfusion for 30 min in the absence (A and B) or presence (B) of ANP, 24-h ischemia in cold UW solution, and reperfusion for 45 min, livers were snap-frozen, homogenized and fractionated as described in “Materials and methods“. Samples were taken at indicated times and total Hsp27 in cytoskeletal (Csk) and cytosolic (Csl) fraction was detected via Western blot analysis using an anti-Hsp27 antibody. One representative Western blot is shown for each treatment group. (A) Effect of ischemia and reperfusion on the distribution of total Hsp27 in the liver. Quantification of total Hsp27 in the liver samples was verified via densitometric analysis of the corresponding bands, which was used to evaluate the ratio of Hsp27 in the cytoskeletal fraction (Csk) resp. cytosolic fraction (Csl) referring to total Hsp27 signal (cytoskeleton+cytosol) in the respective sample. Data are expressed as percent of total Hsp27 in cytoskeletal (F-actin) and cytosolic (G-actin) fraction as mean±SE of n=4 experiments in each treatment group. ^aP<0.05, vs Hsp27 content in cytoskeletal fraction in untreated livers after 30-min perfusion. (B) Distribution of total Hsp27 in untreated (control) and ANP-preconditioned livers after 24-h ischemia. Results are shown as percent of total Hsp27 in cytosolic and cytoskeletal fraction. Data are presented as mean±SE of n=4 experiments in each treatment group. ^cP<0.001 vs Hsp27 content in cytoskeletal fraction of untreated livers after 24-h ischemia. In ANP treated livers the differences in the values were not significant.