Evaluation of matrix metalloproteinases and their endogenous tissue inhibitors in biliary atresia-associated liver fibrosis.

BACKGROUND/PURPOSE: Matrix metalloproteinases (MMPs) and their endogenous tissue inhibitors (TIMPs) are major proteases responsible for remodeling the liver tissue, but their roles in biliary atresia (BA)--associated liver fibrosis are not clear.

METHODS

A DNA microarray containing complementary DNA clones of 10 MMPs and 4 TIMPs was used to compare the expression profiles of the liver cytokines among 3 patients with BA at the time of Kasai procedure (KP) with 3 at the time of liver transplantation (LT). Liver samples from 2 children without liver fibrosis were used as normal controls. Those genes that were differentially expressed by more than 2-fold between groups were further quantified with real time quantitative reverse transcription-polymerase chain reaction (QRT-PCR) and validated with gel electrophoresis.

RESULTS

In normal human liver, messenger RNAs (mRNAs) of TIMP-1, -2, and -3, but not of TIMP-4 and none of the 10 MMPs studied, were expressed in DNA microarray. With progression of liver fibrosis, only mRNA of MMP-7, but not other MMPs, was induced to express at a significantly higher level in the array. Despite its low level of expression, MMP-9 mRNA was significantly upregulated in KP but downregulated in LT, whereas MMP-2, which was not showed in the array, was significantly upregulated in LT than in KP and control in real time QRT-PCR. There was a more than 2-fold increase in TIMP-1 and TIMP-2 mRNA expression in LT over control in the array, which was confirmed in subsequent real time QRT-PCR. The expression of TIMP-3 mRNA was significantly downregulated in KP than in control.

CONCLUSIONS

This study verified differential expression of MMPs and TIMPs in different stages of BA, with emphasis on the role of TIMP-1, -2, and -3 as well as MMP-2, -7, and -9 transcripts in remodeling of liver tissue during the progress of BA-associated liver fibrosis.