Protection of Veratrum nigrum L. var. ussuriense Nakai alkaloids against ischemia-reperfusion injury of the rat liver

Figure 1 Changes of histology in the liver tissue 90 min after ischemia and 240 min after reperfusion in rats (× 400). Five μm sections of liver tissue were stained with hematoxylin and eosin according to standard procedures. For all groups, n = 10. A: Control group: Normal appearance of hepatocytes and sinusoids; B: I/R group: Histological edema, hemorrhage, partial exfoliation of blood vessel endothelium and infiltration with inflammatory cells; C: I/R + VnA (10 μg/kg) group: A significant amelioration of histological edema, hemorrhage, and exfoliation of blood vessel endothelium; D: I/R + VnA (20 μg/kg) group: Slight inflammatory cell infiltration with most hepatocytes in normal appearance.

Figure 2 Plasma aminotransferase (ALT) (A) and acetic dehydrogenase (LDH) (B) levels in different groups (mean ± SD, n = 10). After 90 min of hepatic ischemia and 4 h of reperfusion, plasma levels of ALT and LDH were determined with an OLYMPUS AU5400 automatic analyzer. ^bP < 0.01 vs control group; ^dP < 0.01 vs I/R group.

Figure 3 Activity of superoxide dismutase (SOD) in the liver tissue in different groups (mean ± SD, n = 10). After 90 min of ischemia and 4 h of reperfusion, the liver tissue was homogenized and assayed for SOD levels with an SOD assay kit as the index of hepatic oxidative stress. ^bP < 0.01 vs control group; ^cP < 0.05 vs I/R group.

Figure 4 Activity of myeloperoxidase (MPO) in the liver tissue in different groups (mean ± SD, n = 10). MPO contents in liver tissue were analyzed with an MPO assay kit as the index of neutrophil recruitment. ^bP < 0.01 vs control group; ^dP < 0.01 vs I/R group.

Figure 5 Content of nitric oxide (NO) in the liver tissue in different groups (mean ± SD, n = 10). After 90 min of ischemia and 4 h of reperfusion, the liver tissue was homogenized and assayed for NO levels with an NO assay kit as the index of hepatic oxidative stress. ^bP < 0.01 vs control group. ^aP < 0.05, ^dP < 0.01 vs I/R group.

Figure 6 Immunohistochemical staining of adhesion molecule-1 (ICAM-1) expression in the liver tissue after 90 min of ischemia and reperfusion for 240 min in rats (× 400). Formalin-fixed, paraffin-embedded liver specimens were stained by streptavidin/peroxidase immunohistochemistry technique. For all groups, n = 10. A: Control group; B: I/R group; C: I/R + VnA (10 μg/kg) group; D: I/R + VnA (20 μg/kg) group.

Figure 7 Immunohistochemical staining of E-selectin in liver tissue after ischemia for 90 min and reperfusion for 240 min in rats (× 400). Formalin-fixed, paraffin-embedded liver specimens were stained by the streptavidin/peroxidase immunohistochemistry technique. For all groups, n = 10. A: Control group; B: I/R group; C: I/R + VnA (10 μg/kg) group; D: I/R + VnA (20 μg/kg) group.

Figure 8 Immunohistochemical results (semi-quantitative analysis) of adhesion molecule-1 (ICAM-1) (A) and E-selectin (B) in liver tissue in different groups. Data were presented as mean ± SD. ^bP < 0.01 vs Control group; ^aP < 0.05, ^dP < 0.01 vs I/R group.

Figure 9 Liver adhesion molecule-1 (ICAM-1) (A) and E-selectin (ES) (B) protein signals of Western blot analyzed with a gel imaging system (Kodak system EDAS120, Japan). For all groups, n = 10. L1: Control group; L2: I/R group; L3: VnA (10 μg/kg) group; L4: VnA (20 μg/kg) group. Compared with the control group, the signals in I/R group increased and weakened in the VnA pretreated group in a dose-dependent manner.