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Effect of naphthalene acetic acid on proliferation and apoptosis of mice hepatocytes
ObjectiveTo investigate the effect of naphthalene acetic acid (NAA) on proliferation and apoptosis of mice hepatocytes.MethodsFifty mice were randomly divided into five groups: positive control, normal control, low, middle and high dose of NAA groups.Mice in low, middle and high dose of naphthalene acetic acid groups were fed with 200 mg/kg, 1 000 mg/kg and 5 000 mg/kg NAA animal food, respectively.Mice in positive control, normal control groups were fed with usual animal food.After 4 weeks, mice were sacrificed and liver was separated.Three days before scarification, mice in positive control group were given 20 mg/kg ascyclophosphami daily by intraperitoneal injection.The proliferative activity of hepatocytes was detected by MTT assay.The expressions of PCNA protein and Caspase-3 were measured by immunohistochemistry method.ResultsThe proliferative activities of mice hepatocytes in high dose group and normal control group were 0.794±0.019 and 1.055±0.019, respectively.The expressions of PCNA protein and Caspase-3 in high dose group were 11.2% and 37.9%, and those in the normal group were 33.1% and 6.3%, respectively.All differences were significant (P＜0.01).ConclusionHigh dose of NAA acid could significantly inhibit proliferation and increase apoptosis of hepatocytes in mice.
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联合应用实时量RT-PCR和激光显微切割检测单个肝细胞RNA的表达
AIM: The manner in which a cell responds to and influences its environment is ultimately determined by the genes that are expressed. To better understand cellular functions, the isolation of single cells and subsequent quantification of the expressed genes is essential. METHODS: Normal liver tissue was obtained from operation, snapfrozen in liquid nitrogen and sectioned in crystat. Individual hepatocytes were microdissected. RNA was extracted, then reverse transcribed and amplified using real- time quantitative polymerase chain reaction (PCR). RESULTS: Single hepatocytes were dissected by laser beam and catapulted to the microcentrifuge cap which was put above the slide. In this way,cells were collected, RNA was extracted, reverse transcribed to cDNA and used for analysis of RNA expression by realtime quantitative PCR. The amplification results showed that quantitation of the RNA inside the cell was compatible with the number of cells. CONCLUSION: The expression of RNA in single cells can be quantitated successfully by using laser microdissection and real -time PCR. These techniques provide an opportunity to monitor in vivo gene expression levels in single hepatocytes.
