Expression of tubulin folding cofactor B in mouse hepatic ischemia-reperfusion injury

Abstract

The aim of the present study was to investigate the association between tubulin folding cofactor B (TBCB) expression and ischemia-reperfusion injury (IRI) in mice. A total of 48 C57BL/6 mice were randomly divided into a control group (Sham, n=6) and an ischemia-reperfusion group (n=42). The ischemia-reperfusion group was further divided into 6 subgroups as per different times after reperfusion (2, 4, 6, 8, 12 and 24 h), with 7 mice per subgroup. A hepatic IRI model was established in mice by clamping the hepatic hilum. Morphology, serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α), and the expression level of TBCB were detected. Compared with the control group, the livers from the ischemia-reperfusion group were significantly changed, particularly at 12 h following ischemia-reperfusion, with obvious hepatic cell degeneration and necrosis. The ALT, AST, IL-6 and TNF-α levels in the sera of the mice in the hepatic ischemia-reperfusion group were increased at all time points following ischemia-reperfusion, and were the highest at 12 h, demonstrating statistically significant differences when compared with the control group (P<0.05). Furthermore, the expression levels of TBCB, TNF-α and IL-6 were significantly increased at all time-points following ischemia-reperfusion, and were the most significant at 12 h. At 24 h following ischemia-reperfusion, the expression levels had decreased. The present study indicated that TBCB expression is associated with TNF-α and IL-6 expression levels in mice with hepatic ischemia-reperfusion, and may be key in the development of liver injury during ischemia-reperfusion in mice.

Keywords: cell microtubule; cytoskeleton; ischemia-reperfusion injury; tubulin folding cofactor B.

Figure 1.

Hepatic ischemia-reperfusion injury model. (A-G) Hematoxylin and eosin staining was performed to observe morphological changes in the (A) sham surgery group and (B-G) ischemia-reperfusion groups.

Figure 2.

ALT, AST, IL-6 and TNF-α expression levels were upregulated in hepatic ischemia-reperfusion injury. (A) ALT and (B) AST expression levels in the serum were measured by automatic biochemical analyzer. (C) IL-6 and (D) TNF-α expression levels in the serum were evaluated by enzyme-linked immunosorbent assay. *P<0.05 vs. sham; ^#P<0.05 vs. 12 h. ALT, alanine aminotransferase; AST, aspartate aminotransferase; IL-6, interleukin 6; TNF-α, tumor necrosis factor-α.

Figure 3.

TBCB expression level was upregulated in hepatic ischemia-reperfusion injury. (A) TBCB mRNA expression was analyzed by reverse transcription-quantitative polymerase chain reaction following ischemia-reperfusion. (B) TBCB protein expression was analyzed by western blotting following ischemia-reperfusion. (C) The band intensities were quantified and normalized to β-actin. *P<0.05 vs. sham; ^#P<0.05 vs. 12 h. TBCB, tubulin folding cofactor B.

Figure 4.

TBCB expression level positively correlates with the expression levels of IL-6 and TNF-α. (A-C) Expression levels of IL-6, TNF-α and TBCB in hepatic cells at 6, 8, 12 and 24 h following ischemia-reperfusion were detected by immunohistochemistry. Correlation analysis was performed between (D) TBCB and IL-6 expression levels and (E) TBCB and TNF-α expression levels. TBCB, tubulin folding cofactor B; IL-6, interleukin 6; TNF-α, tumornecrosis factor-α.