动物实验,金属硫蛋白,肝细胞癌 z-bo 2020-08-26 495

　　OBJECTIVE: To understand the dynamic changes and rules of metallothionein (MT) gene expression levels in the occurrence of hepatocellular carcinoma (HCC) in mice, and to explore the importance of MT in the occurrence of liver cancer.

　　Method: 125 male C57BL/6J mice aged 5-8 weeks were randomly divided into normal control group and HCC model group. Mice in the model group were injected intraperitoneally with diethylnitrosamine (100 mg/kg, 50 mg/kg) and ethanol (53% by gavage from the third week, 5 mL/kg, 5 days). /week). , Up to 35 weeks; the normal group was given the same amount of sterile tap water by gavage. Mice were sacrificed on the weekends of experiments 1, 3, 9, 13, 24, and 35, and liver tissue samples were collected to calculate liver index. Histopathology HE, Masson and reticular fiber staining are used to detect liver tissue damage and HCC; enzyme-linked immunosorbent assay is used to detect malondialdehyde (MDA) in liver tissue homogenate. Detect activity; use real-time fluorescent quantitative PCR to analyze the MT transcription level.

　　Result: Progressive liver damage was observed in the mouse model group. At the end of the 35th week, the texture of the liver became significantly harder, and nodules of various sizes appeared on the surface. Approximately 50% of mice have abnormal mitotic patterns and abnormal liver plates. The pathological changes of HCC, such as structure, liver index significantly increased, MDA activity increased at different time periods, MTs mRNA levels significantly increased at various time periods 13 weeks before the experiment, and graded after 24 weeks. Liver fibrosis III was significantly reduced, and the level of MTs mRNA was lower than normal.

　　Conclusion: The level of MTs mRNA in the liver tissue of mice increased significantly from the early stage of injury to the obvious low expression after fibrosis, which indicates that the down-regulation of MTs expression is related to the occurrence of HCC. Being displayed.