Biochemical and Biophysical Research Communications
Transgenic mice expressing osteopontin in hepatocytes as a model of autoimmune hepatitis☆
Section snippets
Materials and methods
Construction of osteopontin transgene. Mouse osteopontin cDNA (0.9 kb) was isolated by RT-polymerase chain reaction (PCR) using total RNA prepared from concanavalin-A-activated spleen cells of C57BL/6 mouse (Japan SLC, Hamamatsu, Japan). HindIII site in the open reading frame of mouse osteopontin cDNA was abolished by silent mutation using a standard PCR method. The mutated osteopontin cDNA was then cloned into the EcoRI site of pLG1-SAP vector [11] that contains human serum amyloid P component
Transgenic mouse lines expressing osteopontin
Among 52 mice brought up after the transplantation, 6 mice (3 males and 3 females) carried the transgene. These 6 mice were mated with C57BL/6 mice at 8 weeks of age, and the transgene was transmitted to the offspring of 4 lines; the SAP-osteopontin 17, 35, 43, and 73 lines. The transgenic mice were mated each other in the same lines, and the offspring carrying the transgene derived from the SAP-osteopontin 17, 43, and 73 lines were used in the experiments. The offspring from the
Discussion
In the present study, four lines of osteopontin transgenic mice were obtained. We used a pLG1-SAP vector [11] that contains the human SAP promoter consisting of a motif putatively recognized by hepatocyte nuclear factor (HNF)-1 [14] and the exon and intron of rabbit β-globin gene, because osteopontin may be expressed exclusively in hepatocytes in such transgenic mice. In fact, Western blot analysis using isolated liver cells and immunohistochemical examination revealed that osteopontin protein
Acknowledgements
We thank Ms. Kayoko Naiki (Department of Internal Medicine, Saitama Medical School) for her technical help.
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Mechanism of hepatitis C virus (HCV)-induced osteopontin and its role in epithelial to mesenchymal transition of hepatocytes
2013, Journal of Biological ChemistryA humanized anti-osteopontin antibody protects from Concanavalin A induced-liver injury in mice
2011, European Journal of PharmacologyCitation Excerpt :Accumulative evidences demonstrated the role of osteopontin in the development of immune-mediated hepatitis. Transgenic mice expressing osteopontin in hepatocytes also showed significant autoimmune hepatitis (Mochida et al., 2004), and T cell mediated-liver injury was suppressed in osteopontin-deficiency mice (Diao et al., 2004). A murine anti-human osteopontin monoclonal antibody (23C3 mAb), which cross-reacted with murine osteopontin, was able to inhibit lymphocyte migration, and reduce the severity of established arthritis in mice (Fan et al., 2008).
Transgenic expression of osteoactivin in the liver attenuates hepatic fibrosis in rats
2007, Biochemical and Biophysical Research CommunicationsCitation Excerpt :In this study, we provide the first direct evidence that transgenic expression of OA in the liver inhibited hepatic fibrosis in rats fed a CDAA diet for 12 weeks. Transgenic expression of OA was driven by the SAP promoter, which induces specific gene expression in hepatocytes, but not non-parenchymal cells [17]. Although transgenic expression of OA was only induced by the SAP promoter at low levels, hepatocyte expression of OA in OA-Tg rats was able to reduce hepatic fibrosis in association with a decrease in the number of activated HSCs.
Plasma osteopontin levels in patients with fulminant hepatitis
2004, Hepatology Research
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Grant-in-Aid for scientific research by Japan Society for the Promotion of Science (JSPS), Scientific Research (C) (2): 14670505.