SPECT imaging was performed in control rats and TAA-treated rats (n = 3 per group). Each animal was administered 6 μCi of 99mTc-cRGD by way of the penile vein. Animals were placed supine on a SPECT meter (Philips IRIX,

Best, Netherlands). Anterior images were acquired 15, 30, and 45 minutes after the injection and stored digitally. Then a computer-aided manipulator discriminated the region of interest in the liver and heart and the radioactivity ratio (counts/pixel) of liver to heart was calculated. A tracer selleck products dose (6 μCi) of 125I-cRGD was intravenously administrated to control rats and TAA-treated rats (n = 3 per group). Additionally, 6 μCi 125I-cRGD was also administered simultaneously with excessive unlabeled cRGD (500-fold high dosage of 125I-cRGD) (n = 3 per group). Blood samples were collected by heart puncture 45 minutes after dosage and the organs and tissues were collected, washed in saline, and weighed. Subsequently, radioactivity in the samples was determined by a gamma-counter. The total radioactivity per organ was calculated and corrected for the blood-derived radioactivity. The organ Selleckchem PF 2341066 accumulation of 125I-cRGD was calculated as a percentage of the injected dose per gram

of wet tissue mass (%ID/g). All collected data were expressed as mean ± standard deviation (SD). Comparisons between groups were achieved by one-way analysis of variance tests (ANOVA) followed by post-hoc tests with SPSS 11.5 statistical software (Chicago, IL) and P < 0.05 was considered statistically significant. The purity of cRGD and all of its derivatives was above 95%. The molecular weight is 693 for cRGD and 962.01 for FAM-cRGD. Hepatocyte injury and fibrotic septa formation were observed

in the livers of TAA-3w rats and the fibrotic area was 5.8 ± 1.2% (Ishake score 1.8 ± 0.6, representing as mild fibrosis). In the livers of TAA-9w rats, extensive bridging fibrosis in addition to a distortion of liver architecture with pseudo-lobule formation was visible. The fibrotic area was significantly increased to 16.5 ± 3.6% (Ishake score 5.3 ± 0.7, representing MCE as advanced fibrosis) (P < 0.05) (Fig. 1A,B). Hydroxyproline content in liver tissue was markedly increased with the progression of liver fibrosis (Fig. 1C). Serum ALT and AST levels in the TAA-3w group was higher than in the control group and the TAA-9w group (P < 0.05 for all comparison) (Fig. 1D). With the progression of liver fibrosis, hepatic mRNA levels and protein levels of αv and β3 integrin subunits and α-SMA were markedly increased and were the highest in rats with advanced fibrosis (Fig. 1E,F). To colocalize expression of integrin αvβ3 with albumin, α-SMA, CD31, CD68, and CD163, double immunofluorescent staining was performed in the livers of advanced fibrosis. As shown in Figs. 2, 3, the positive staining of integrin αvβ3 was mainly overlapped with α-SMA staining (Fig. 2B).