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Interleukin‐1β induces tumor necrosis factor‐α secretion from rat hepatocytes
Author(s) -
Yoshigai Emi,
Hara Takafumi,
Inaba Hiroyuki,
Hashimoto Iwao,
Tanaka Yoshito,
Kaibori Masaki,
Kimura Tominori,
Okumura Tadayoshi,
Kwon AHon,
Nishizawa Mikio
Publication year - 2014
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.12157
Subject(s) - tumor necrosis factor alpha , rna , lipopolysaccharide , messenger rna , biology , cytokine , microbiology and biotechnology , oligonucleotide , interleukin , northern blot , immunology , gene , biochemistry
Aim Tumor necrosis factor‐α ( TNF ‐α) is a pleiotropic cytokine involved in various inflammatory diseases. The only production of TNF ‐α in the liver is thought to be from hepatic macrophages known as K upffer cells, predominantly in response to bacterial lipopolysaccharide ( LPS ). Methods Primary cultured rat hepatocytes were used to analyze TNF ‐α expression in response to the pro‐inflammatory cytokine, interleukin‐1β ( IL ‐1β). Livers of rats subjected to LPS ‐induced endotoxemia were analyzed. Results Immunocytochemistry and enzyme‐linked immunosorbent assays demonstrated that IL ‐1β‐treated rat hepatocytes secreted TNF ‐α, and RNA analyses indicated that TNF ‐α m RNA was induced specifically by IL ‐1β. Northern blot analysis showed that not only mRNA , but also a natural antisense transcript (as RNA ), was transcribed from the rat T nf gene in IL ‐1β‐treated hepatocytes. TNF ‐α was detected in the hepatocytes of LPS ‐treated rats. Both TNF ‐α m RNA and as RNA were expressed in the hepatocytes of LPS ‐treated rats, human hepatocellular carcinoma and human monocyte/macrophage cells. To disrupt the interaction between TNF ‐α as RNA and TNF ‐α m RNA , sense oligonucleotides corresponding to TNF ‐α m RNA were introduced into rat hepatocytes resulting in significantly increased levels of TNF ‐α mRNA . One of these sense oligonucleotides increased a half‐life of TNF ‐α m RNA , suggesting that the TNF ‐α as RNA may reduce the stability of TNF ‐α m RNA . Conclusion IL ‐1β‐stimulated rat hepatocytes are a newly identified source of TNF ‐α in the liver. TNF ‐α m RNA and as RNA are expressed in rats and humans, and the TNF ‐α as RNA reduces the stability of the TNF ‐α m RNA . Hepatocytes and TNF ‐α as RNA may be therapeutic targets to regulate levels of TNF ‐α m RNA .