Table 1.
Effect of ELAP on body weight, liver weight, and liver index after 8 weeks.
Figure 1.
(A) Normal control liver has regular smooth surface. (B) TAA control liver has rough nodular surface, with uniform distribution of micronodules (<0.3 cm) and macronodules (≥0.3 cm). (C) TAA+silymarin liver has normal smooth surface. (D) TAA+250 mg/kg ELAP liver has nearly smooth surface and few micronodules. (E) TAA+500 mg/kg ELAP liver has normal smooth surface and nearly normal anatomical shape and appearance. Livers shown are representative samples (n = 6/group).
Figure 2.
H&E staining of histopathological liver sections.
(A) Normal control liver has normal histological structure and architecture. (B) TAA control liver has structural damage, irregular regenerating pseudolobules with dense fibrotic septa, proliferation of bile duct, and presence of centrilobular and inflammatory cells. (C) TAA+silymarin liver has mild inflammation but no fibrotic septa. (D) TAA+250 mg/kg ELAP liver has partially preserved hepatocytes, small area of necrosis, narrow fibrotic septa. (E) TAA+500 mg/kg ELAP liver has partially preserved hepatocytes and small areas of mild necrosis. Sections shown are from representative samples (n = 6/group).
Figure 3.
Masson’s Trichrome staining of histopathological liver sections.
(A) Normal control liver has normal architecture. (B) TAA control liver shows proliferation of bile duct, dense fibrous septa, and collagen fibers. (C) TAA+silymarin liver shows minimal fibrous septa and collagen fibers. (D) TAA+250 mg/kg ELAP liver shows moderate fibrous septa and irregular regenerating nodules. (E) TAA+500 mg/kg ELAP liver shows mild fibrous septa and collagen fibers. [Sections shown are from representative samples (n = 6/group)].
Figure 4.
PCNA labeling in histopathological liver sections.
(A) Normal controls stained without primary antibody show normal liver architecture and no PCNA labeling. (B) TAA controls have many PCNA-positive hepatocyte nuclei. (C) TAA+silymarin rats have no PCNA-positive hepatocytes. (D) TAA+250 mg/kg ELAP rats have moderate hepatocyte regeneration, as indicated by moderate presence of PCNA-positive hepatocyte nuclei. (E) TAA+500 mg/kg ELAP rats have mild PCNA expression with few regenerative hepatocytes.
Table 2.
Effect of ELAP on PCNA labeling and mitotic index.
Figure 5.
Effect of ELAP on SOD activity in liver tissue.
Data expressed as mean ± SEM (n = 6 rats/group). *P<0.05 compared with TAA control, #P<0.05 compared with normal control.
Figure 6.
Effect of ELAP on CAT activity in liver tissue.
Data expressed as mean ± SEM (n = 6 rats/group). *P<0.05 compared with TAA control, #P<0.05 compared with normal control.
Figure 7.
Effect of ELAP on MDA levels in liver tissue.
Data expressed as mean ± SEM (n = 6 rats/group). *P<0.05 compared with TAA control, #P<0.05 compared with normal control.
Figure 8.
Effect of ELAP on NO levels in liver tissue.
Data expressed as mean ± SEM (n = 6 rats/group). *P<0.05 compared with TAA control, #P<0.05 compared with normal control.
Table 3.
Effect of ELAP on biochemical parameters in TAA-induced liver cirrhosis.
Table 4.
Effect of ELAP on serum liver biomarkers in TAA-induced liver cirrhosis.
Table 5.
IC50 values of isolated fractions on HepG2 and WRL-68 cells after 24 h.
Figure 9.
Effect of ELAP fractions on LDH leakage in HepG2 cells.
LDH assay was used to assess the loss of membrane integrity in HepG2 cells treated with CF, HF, or BF. Significant cytotoxicity was observed at 12.5 to 100 µg/mL. Data represent mean ± SD of three independent experiments. *P<0.05 compared with no treatment.
Figure 10.
Effect of ELAP fractions on ROS generation in HepG2 cells.
The level of ROS increased significantly after CF and HF treatment at concentrations from 25 to 100 µg/mL. Data represent mean ± SD of three independent experiments. *P<0.05 compared with no treatment.
Figure 11.
Effect of ELAP fractions on apoptotic markers in HepG2 cells.
(A) HepG2 cells were treated with medium alone and 12.5 µg/mL CF, HF, or BF, then stained with Hoechst 33342, cell membrane permeability, MMP, and cytochrome c dyes. Isolated fractions caused a marked elevation in cytochrome c release and cell membrane permeability, and a noticeable decrease in MMP. (B) Dose-dependent reduction of MMP and increase in cell permeability in treated HepG2 cells (12.5 to 100 µg/mL). Cytochrome c was significantly released at CF, HF, or BF concentrations of 12.5 to 100 µg/mL. Data represent mean ± SD of three independent experiments. *P<0.05 compared with no treatment.
Figure 12.
Effect of ELAP fractions on Caspases activation in HepG2 cells.
Relative luminescence dose-dependent expression of caspase-3/7, -8, and -9 in HepG2 cells treated with CF and HF caused significant activation at 12.5 to 100 µg/mL concentrations. In HepG2 cells treated with CF and HF, caspase-3/7, -8, and -9 expression increased significantly and in a dose-dependent manner at concentrations from 12.5 to 100 µg/mL. BF activated caspase-3/7 and -8 at the same concentrations, but caspase-9 was activated only at higher concentrations (50 and 100 µg/mL). Activation was measured by relative luminescence. Data represent mean ± SD of three independent experiments. *P<0.05 compared with no treatment.