Selection of reference genes for quantitative real-time reverse transcription-polymerase chain reaction in concanavalin A-induced hepatitis model
Section snippets
Animal treatment
C57BL/6 male mice, 8–10 weeks of age, were purchased from Shanghai Laboratory Animal Company (SLAC, Shanghai, China). Mice were housed in the animal facilities of the Shanghai Institute of Endocrine and Metabolic Diseases, Shanghai Jiao-Tong University School of Medicine, under pathogen-free conditions according to the institutional animal care and use committee guidelines. Mice were fed ad libitum a standard laboratory chow diet provided by SLAC.
CIH model
Mice (4 or 5 per group) were injected via the
ConA treatment induces hepatitis and hepatocellular apoptosis
Hepatic changes were examined histologically and biochemically at different time points after ConA injection. Consistent with a previous report [23], serious necrotic phenotypes were observed in livers at 8 and 24 h after ConA injection, whereas the necrotic phenotype nearly disappeared at 72 h (Fig. 1A). Simultaneously, based on the genomic DNA fragmentation analysis [16], hepatocytes demonstrated obvious apoptosis at 8 h as compared with the control group, and even more serious at 24 h,
Discussion
Hepatitis is a potentially life-threatening liver disease. Mouse hepatitis models provide valuable insights in attenuating and curing the disease. CIH in mouse is a model developed during recent years and is used with increasing interest to investigators of liver disease [32]. Gene expression profiles at mRNA and protein levels during hepatitis progression are extremely valuable for functional analysis. Q-PCR analysis provides great convenience and accuracy in determination of target gene mRNA
Acknowledgments
This work was supported by the National High Technology Research and Development Program of China (2006AA02A409) and the National Natural Science Foundation of China (30973571).
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