Figure 1.
Illustrations of surgical procedure of PVL+ISS.
(A) Schematic anatomy of liver lobes and portal veins: right median lobe (RML), left median lobe (LML), left lateral lobe (LLL), right lobe (RL), caudate lobe (CL). The silk knots represent the ligated lobes. (B) Portal veins (PV) of rats in surgery. (C) Ischemic line emerged to the right of the falciform ligament when the left branch of the portal vein was ligated. (D) The median lobe was in situ split along the ischemic line.
Table 1.
Primer sequences.
Figure 2.
The hepatic regeneration rate (HRR) of the right median lobe after SHAM, PVL and PVL+ISS.
Values are means±SD. The HRR of PVL or PVL+ISS was significantly different than the SHAM group at all time points, and the HRR of PVL+ISS was significantly different compared with the PVL group at 72 h and 7 days, but no obvious difference was observed at 24 and 48 h (#P<0.01 compared with the sham group; *P<0.01 compared with the PVL group).
Figure 3.
The number of Ki-67-positive hepatocytes in regenerating lobes after PVL and PVL+ISS.
(A) Immunohistochemical staining for Ki-67 in regenerating lobes at all time points. Hepatocytes with nuclear deposition of brownish pigment were considered positive (original magnification 200×). (B) The regenerating lobes were analyzed for Ki-67-positive hepatocytes at all time points. The number of positive hepatocytes per visual field (200×) is presented. The increase in Ki-67-positive cells shows a significant difference at 48 and 72 h (*p<0.01 and *p<0.05, respectively), while no significant difference was observed at 24 h and 7 days.
Figure 4.
Laser speckle contrast imaging (LSCI).
Typical hepatic flux perfusion images obtained from LSCI of the RML and the LML are shown: 5 min after laparotomy (A), 5 min after PVL (B) and 5 min after PVL+ISS (C). The microcirculation of the right median and left median lobes measured by LSCI after PVL and PVL+ISS was expressed as a percentage of the baseline value (D). There was a significant difference in the left median lobe before and after ISS (*p<0.01), but no significant difference was observed in the right median lobe (p>0.05).
Figure 5.
Biochemical indications after different surgical interventions at all time points.
Serum levels of ALT (A), AST (B), ALB (C) and TBIL (D). Values are means±SD. There was a significant difference in ALT and AST release between PVL and PVL+ISS at 24 h (*p<0.05) after surgery and between PVL or PVL+ISS and SHAM at 24 and 48 h (#p<0.01). Then, they returned to baseline values at 72 h. No significant difference was observed at 72 h and 7 days in all groups. Hepatocellular synthetic function evaluated by serum albumin level showed a prolonged dysfunction in both PVL and PVL+ISS groups (#P<0.05 compared with the sham group at all time points). The serum albumin level of PVL+ISS was significantly different at 24, 48 and 72 h compared with the PVL group (*p<0.05). The serum levels of TBIL in all groups were not significantly different at any time point.
Figure 6.
H-E staining of left median lobe at 24 h after surgery.
Necrosis in PVL (A) and PVL+ISS (B) at 24 h after surgery. The total necrosis area scores of the left median lobes observed in 10 random visual fields (100×). Values are means±SD. There was a larger necrosis area in PVL+ISS than PVL. The necrosis area of the left median lobe in PVL+ISS was significantly different compared with that of the PVL group at 24 h (*p<0.05) (C).
Figure 7.
The mRNA expression of cytokines in regenerating liver tissue at 24 h after PVL and PVL+ISS.
The relative mRNA expression levels of TNF-α, IL-6, HGF and HSP70 were determined by RT-PCR and given as fold inductions relative to the sham-operated livers. (#P<0.01 compared with the sham group; *P<0.01 compared with the PVL group).
Figure 8.
Protein levels of cytokines in regenerating liver tissue at 24 h after different surgical interventions.
The protein levels of hepatic TNF-α, IL-6 and HGF in the PVL+ISS group had a significant difference compared with the PVL group. Both groups were significantly different from the SHAM group (#P<0.05 compared with the SHAM group; *P<0.01 compared with the PVL group).