Stromal cell derived factor-1 enhances bone marrow mononuclear cell migration in mice with acute liver failure

Abstract

Aim: To evaluate the number of bone marrow mononuclear cells (BMMC) that are migrated to the liver following transplantation of murine BMMC into mice with acute liver injury.

Methods: BMMC were isolated from the bone marrow of mice in a lymphocyte separation medium and then labeled with PKH26. The labeled cells were subsequently infused into the caudal veins of BALB/c mice with hepatic injury induced by carbon tetrachloride and 2-acetylaminofluorene. Mice in experimental group were treated with stromal cell-derived factor-1 (SDF-1) which was injected intraperitoneally after transplantation of BMMC. Mice in control group were injected intraperitoneally with 0.1 mL of saline (0.9% NaCl) after transplantation of BMMC. After 2 wk, migration of the cells in experimental group was studied by fluorescence microscopy. The expression of proliferating cell nuclear antigen and albumin was quantified with manual methods in both groups. The serum transaminase levels at different time points were compared between the two groups.

Results: The labeled "cells" were found in the portal region and central veins of hepatic lobules. The PKH26-labeled cells appeared at an average frequency of 108 +/- 8/high power field in the experiment group and 65 +/- 8/high power field in the control group (P < 0.05). The total number of positive cells was 29 +/- 7/high power field in the experimental group and 13 +/- 2/high power field in the control group. The albumin expression level was also higher in the experimental group than in the control group (29 +/- 7 vs 13 +/- 2, P < 0.05). The total number of crossing points was 156 +/- 5/high power field in the experimental group and 53 +/- 5/high power field in the control group (P < 0.05). The serum alanine aminotransferase levels in experimental and control groups were measured at different time points (120 +/- 40 vs 118.50 +/- 1.75, P > 0.05; 80.60 +/- 6.50 vs 101.08 +/- 5.67, P < 0.05; 50.74 +/- 5.38 vs 80.47 +/- 4.62, P < 0.05; 30.54 +/- 2.70 vs 60.72 +/- 4.37, P < 0.05; 30.77 +/- 5.36 vs 40.47 +/- 6.50, P < 0.05). At the same time, the serum aspartate aminotransferase levels were measured in experimental and control groups at different time points (122.55 +/- 1.46 vs 120.70 +/- 4.22, P > 0.05; 54.26 +/- 6.50 vs 98.70 +/- 8.20, P < 0.05; 39.47 +/- 5.39 vs 78.34 +/- 4.50, P < 0.05; 28.94 +/- 2.70 vs 56.44 +/- 4.28, P < 0.05; 30.77 +/- 5.45 vs 42.50 +/- 6.28, P < 0.05).

Conclusion: SDF-1 can promote the migration of BMMC to the liver of mice with acute liver failure.

Figure 1

Histopathology of hepatic tissue from the two groups. A: PKH26-labeled cells detected after establishment of acute liver failure animal model with extensive vacuolar degeneration and edema of hepatocytes in acute liver failure (A1) and normal liver tissue (A2); B: Sporadic PKH26-labeled bone marrow stem cells in experimental group (B1) and control group (B2); C: Expression of PCNA in sporadic PKH26-labeled bone marrow stem cells in experimental group (C1) and control group (C2); D: Expression of albumin and sporadic PKH26-labeled bone marrow stem cells in experimental group (D1) and control group (D2) ( × 200).

Figure 2

PKH26-labeled cells detected in experimental and control groups. Data are expressed as mean ± SD. ^aP < 0.05 vs experimental group.

Figure 3

Confocal microscopy shows red fluorescence of cell location and green fluorescence of albumin. The red fluorescence cells could be found in liver tissue of recipient mice, suggesting that PHK-26 positive cells can emerge out of the red fluorescence (A1-A4). The albumin expressed in hepatocytes showed green fluorescence (B1-B4). After the red and green fluorescence cells were located, yellow cells were found in a suitable location (C1-C4).

Figure 4

Detection of serum ALT (A) and AST (B) activity in the two groups. The serum ALT and AST levels were measured with an automatic biochemistry analyzer in experimental and control groups. There was a significant difference between the two groups. At the same time, the serum AST level was measured. ^aP < 0.05 vs experimental group.