Molecular mechanism investigation of cycloheximide-induced hepatocyte apoptosis in rat livers by morphological and microarray analysis

• Published in: Toxicology (Amsterdam) 2006-02, Vol.219 (1), p.175-186
• Main Authors: Ito, Kazumi, Kiyosawa, Naoki, Kumagai, Kazuyoshi, Manabe, Sunao, Matsunuma, Naochika, Yamoto, Takashi
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Biological and medical sciences; Cycloheximide; Cycloheximide - toxicity; Cytokines - biosynthesis; DNA Primers; Endoplasmic Reticulum - drug effects; Endoplasmic Reticulum - metabolism; Endoplasmic reticulum stress; Hepatocytes - drug effects; Hepatocytes - pathology; In Situ Nick-End Labeling; Inflammation - pathology; Kupffer Cells - pathology; Kupffer Cells - ultrastructure; Liver; Liver - cytology; Liver - drug effects; Male; Medical sciences; Microarray; Oligonucleotide Array Sequence Analysis; Pesticides, fertilizers and other agrochemicals toxicology; Protein Synthesis Inhibitors - toxicity; Rats; Rats, Inbred F344; Reverse Transcriptase Polymerase Chain Reaction; Toxicology; Transcription Factors - drug effects; Transcription Factors - metabolism
• ISSN: 0300-483X; 1879-3185
• DOI: 10.1016/j.tox.2005.11.017

Male F344 rats were intravenously treated with 6 mg/kg cycloheximide (CHX), and microarray analysis was conducted on their livers 1, 2 and 6 h after the CHX treatment. The histopathological examination and serum chemistry results indicated a mild hepatic cell death 2 and 6 h after the CHX treatment, respectively. Multi-focal hepatocellular necrosis with slight neutrophil infiltration was observed 6 h after the CHX treatment. The TUNEL staining results showed that the number of apoptotic hepatocytes was the highest 2 h after the CHX treatment. Dramatic increases in the mRNA levels of ATF3 and CHOP genes, both of which were reported to play roles in the ER stress-mediated apoptosis pathway, were observed from 1 h after the CHX treatment. In addition, increase of GADD45, p21 and p53 mRNA levels also suggested a time course-related stimulation of hepatocellular apoptotic signals. These results suggest that the hepatocyte apoptosis induced by the CHX treatment is triggered by ER stress. The hepatic mRNA levels of proinflammatory genes, such as TNFα, IL-1α and β, were also increased 1 and 2 h after the CHX treatment, supposedly mediated by the activated Kupffer cells engulfing the apoptotic hepatocytes.