Fig 1.
Histological analysis of hamster liver.
(A) Microphotographs of Opisthorchis felineus infected hamster liver at 8 weeks post-infection. An adult fluke is found in bile duct lumen. Small bile duct proliferation and chronic cholangitis accompanied by inflammatory infiltration in the bile duct wall; hematoxylin-eosin staining, ×100. (B) Mild portal fibrosis with thin septa formation (portal to portal and portal to central) and (C) pronounced periductal fibrosis with chronic inflammation around the bile duct are observed. In adjacent hepatic tissue, fibrosis with porto-portal septa formation takes place. Van Gieson’s staining showing extensive deposition of mature collagen fibers, ×100. (D) Microphotographs of the uninfected hamster liver. No pathological changes are observed. Hematoxylin-eosin staining, ×100 and (E) Van Gieson’s staining, ×100.
Table 1.
Liver tissue biochemical analysis.
Fig 2.
T2-weighted (A) axial and (B, C) coronal images of O. felineus infected liver and (D) axial and (E, F) coronal images of uninfected hamster liver (control) at 11.7 T. TurboRARE (Rapid Imaging with Refocused Echoes) with respiratory triggering, with pulse sequence parameters TEeff = 18 ms and TR = 900 ms, Flip Angle = 180°, RARE Factor = 4. Bruker, Biospec 117/16 USR, Germany. In O. felineus infected hamster liver, the hyperintense signals from the bile duct wall as well as the bright zones extending from the dilated bile duct due to inflammation and fibrosis are clearly depicted. There are no pathological changes in the liver of the control hamsters.
Fig 3.
Typical in vivo 1H magnetic resonance spectrum of hamster liver.
Spectrum was recorded using the spatially localized PRESS (Point Resolved Spectroscopy) method with TE = 20 ms, TR = 2.5 ms, 256 replicates, voxel size 5×5×5 mm. Water signal in the spectra was suppressed by the variable power and optimized relaxation delays (VAPOR) method. Bruker, Biospec 117/16 USR, Germany. Lipid resonance peaks are labeled: P1: methyl proton CH3 (0.9 ppm); P2: methylene (–CH2–)n (1.3 ppm); P3: -CH2-CH = CH- (2.0 ppm); P4: -CH2-COO- (2.2 ppm); P5: -CH = CH-CH2-CH = CH- (2.8 ppm).
Table 2.
Levels of hepatic lipid metabolites (in vivo 1H MRS).
Fig 4.
Typical in vivo hepatic 31P magnetic resonance spectrum of the right upper abdominal quadrant (includes liver, abdominal wall, subcutis, etc.) of a hamster.
Spectrum was recorded using the nonspatially localized, single-pulse method with TE = 3 ms, TR = 100 ms, 1048 replicates. Bruker, Biospec 117/16 USR, Germany. Phosphorylated metabolites resonance peaks are labeled: P1: phosphomonoester (PME); P2: inorganic phosphate (Pi); P3: phosphodiester (PDE); P4: creatine phosphate (PCr); P5: NTP-γ; P6: NTP-α; P7: NTP-β.
Table 3.
Levels of hepatic phosphorylated metabolites (in vivo 31P MRS).
Fig 5.
Analysis of the AMPK phosphorylation level in the hamster liver.
(A) Western blots of (p-AMPK) phospho-AMPK and (b-actin) beta-actin in O. felineus infected and healthy (control) hamster liver; (M) Precision Plus Protein Western C Standards (BioRad) with Precision Protein StrepTactin-HRP Conjugate (BioRad) are used as a molecular weight marker. (B) The histogram depicts the level of phospho-AMPK determined through densitometry relative to beta-actin band. The values are represented as means±SD (n = 8 animals per group), *p<0.05 denotes significant differences between the infected and the control groups.