Premium
Establishment of mouse Mac‐2 binding protein enzyme‐linked immunosorbent assay and its application for mouse chronic liver disease models
Author(s) -
Iwata Ayumi,
Kamada Yoshihiro,
Ebisutani Yusuke,
Yamamoto Akiko,
Ueda Yui,
Arai Hitomi,
Fujii Hironobu,
Takamatsu Shinji,
Maruyama Nobuhiro,
Maeda Masahiro,
Takehara Tetsuo,
Miyoshi Eiji
Publication year - 2017
Publication title -
hepatology research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.123
H-Index - 75
eISSN - 1872-034X
pISSN - 1386-6346
DOI - 10.1111/hepr.12819
Subject(s) - medicine , fibrosis , steatohepatitis , chronic liver disease , nonalcoholic fatty liver disease , fatty liver , liver disease , biomarker , antibody , monoclonal antibody , pathology , disease , gastroenterology , biology , immunology , cirrhosis , biochemistry
Aim We identified Mac‐2 (galectin‐3) binding protein (Mac‐2bp) as a novel diagnostic and liver fibrosis predicting biomarker for nonalcoholic steatohepatitis in humans. In mouse models, there are no serum biomarkers predicting liver disease severity. In this study, we developed a mouse Mac‐2bp enzyme‐linked immunosorbent assay (ELISA) system and determined its efficacy for predicting the severity of liver disease in mouse models, especially in non‐alcoholic fatty liver disease (NAFLD) models. Methods We established several rat monoclonal antibodies against mouse Mac‐2bp, selected two clones for the ELISA, and checked the accuracy and reproducibility of the ELISA, especially for NAFLD models and liver fibrosis models. We also investigated the relationships between serum levels and hepatic gene expression of Mac‐2bp in mouse models. Results Our ELISA system had high accuracy and reproducibility (R 2 = 0.999). The intra‐assay and inter‐assay results for the coefficient of variation were 2.0–3.7% and 1.7–6.9%, respectively. The levels of bilirubin, hemoglobin, and chyle did not affect the Mac‐2bp serum levels detected by our ELISA kit. In the mouse models, serum Mac‐2bp levels increased with liver disease progression (F0/F1/F2/F3, 239.1 ± 36.7 / 259.1 ± 43.0 / 457.5 ± 162.0 / 643.7 ± 116.0 ng/mL; P < 0.0001), and were significantly correlated with hepatic gene expression of Mac‐2bp ( R = 0.42, P < 0.0001). Conclusion Our mouse Mac‐2bp ELISA system effectively predicts severity of NAFLD and liver fibrosis in mouse models.