C57BL/6J小鼠,肝细胞癌,金属硫蛋白 z-bo 2020-11-13 378

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

　　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 intraperitoneally injected with diethylnitrosamine (100 mg/kg, 50 mg/kg) in the first and second weeks, and were given ethanol (53%, 5 mL/kg, 5 mL/kg) by gavage from the third week. d/week), until 35 weeks; the normal group was given the same amount of sterilized tap water by gavage. Mice were sacrificed on the weekend of experiment 1, 3, 9, 13, 24, and 35, and liver tissue samples were collected and liver index was calculated. Histopathological HE, Masson and reticular fiber staining were used to detect the occurrence of liver tissue damage and HCC; enzyme-linked immunosorbent assay was used to detect the activity of malondialdehyde (MDA) in liver tissue homogenate; real-time fluorescent quantitative PCR was used to analyze the transcription level of MTs .

　　Results: Progressive liver lesions were seen in the model group mice. At the end of the 35th week, the liver texture was significantly hardened, and nodules of different sizes were seen on the surface. About 50% of the mice showed abnormal mitotic patterns of liver cells and abnormal liver plate structure. HCC has pathological changes with significantly increased liver index; increased MDA activity in different periods; significantly increased levels of MTs mRNA in each period before 13 weeks of the experiment, and grade III fibrosis and MTs mRNA levels can be seen after 24 weeks Significant decline, even lower than normal.

　　Conclusion: The level of MTs mRNA in the liver tissue of mice increased significantly from the initial stage of injury to the low expression after significant fibrosis, indicating that the down-regulation of MTs expression is related to the occurrence of HCC.