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Toxicologic Pathology
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Articles

Ferritin Expression in Rat Hepatocytes and Kupffer Cells after Lead Nitrate Treatment

Yang Fan1,4, Toshiyuki Yamada1, Takeshi Shimizu1, Naoki Nanashima1,2, Miki Akita1, Kohji Suto3 and Shigeki Tsuchida1

1 Department of Biochemistry and Genome Biology, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan
2 Department of Medical Technology, Hirosaki University Graduate School of Health Sciences, Hirosaki 036-8564, Japan
3 Department of Internal Medicine, Hirosaki City Hospital, Hirosaki 036-8004, Japan
4 Research Center of Affiliated Shengjing Hospital, China Medical University, Shenyang, 110004, China

Correspondence: Shigeki Tsuchida, Department of Biochemistry and Genome Biology, Hirosaki University Graduate School of Medicine, 5 Zaifu-Cho, Hirosaki 036-8562, Japan; e-mail:tsuchida{at}cc.hirosaki-u.ac.jp.

Lead nitrate induces hepatocyte proliferation and subsequent apoptosis in rat livers. Iron is a constituent of heme and is also required for cell proliferation. In this study, the expression of ferritin light-chain (FTL), the major iron storage protein, was investigated in rat livers after a single intravenous injection of lead nitrate. Western blotting and immunohistochemistry revealed that FTL was increased in hepatocytes around the central veins and strongly expressed in nonparenchymal cells. Some FTL-positive nonparenchymal cells were identified as Kupffer cells that were positive for CD68. FTL-positive Kupffer cells occupied about 60% of CD68-positive cells in the periportal and perivenous areas. The relationships between FTL expression and apoptosis induction or the engulfment of apoptotic cells were examined. TUNEL-positive cells were increased in the treatment group, and enhanced expression of milk fat globule EGF-like 8 was demonstrated in some Kupffer cells and hepatocytes, indicating enhanced apoptosis induction and phagocytosis of apoptotic cells. FTL-positive Kupffer cells were not detected without lead nitrate treatment or in rat livers treated with clofibrate, which induces hepatocyte proliferation but not apoptosis. These results suggest that FTL expression in Kupffer cells after lead treatment is dependent on phagocytosis of apoptotic cells.

Key Words: Lead nitrate • ferritin • cell proliferation • apoptosis • phagocytosis • Kupffer cell

Abbreviations: ABC, avidin-biotin-peroxidase complex • {alpha}-SMA, {alpha}-smooth muscle actin • DAB, 3, 3-diaminobenzidine tetrahydrochloride • GST, glutathione S-transferase • FTL, ferritin light-chain • IRP, iron-regulatory protein • MFG-E8, milk fat globule EGF factor 8 • NPC, nonparenchymal cell • PS, phosphatidylserine • RT-PCR, reverse transcriptase-polymerase chain reaction • SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis • TUNEL, TdT-mediated dUTP-biotin nick end labeling

Toxicologic Pathology, Vol. 37, No. 2, 209-217 (2009)
DOI: 10.1177/0192623308328544
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