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目的 :确定小鼠应用核桃楸青果皮等浸出物后腹腔巨噬细胞内TNFα -mRNA表达的变化。方法 :雌雄小鼠随机分组 ,口服给药后制备巨噬细胞涂片 ;克隆hTNFcDNA的PGEM - 1质粒转化EcoliJM 10 3,选择阳性克隆细胞培养扩增 ;随机引物法制备地高辛标记的探针 ;利用原位杂交技术检测巨噬细胞内TNFα -mR NA的表达。结果 :抗肿瘤核桃楸青果皮等浸出物可增强小鼠腹腔巨噬细胞内TNFα -mRNA表达。结论 :本项目研究揭示了抗肿瘤中草药核桃楸青果皮等浸出物用于抗肿瘤的作用机理。
OBJECTIVE : To determine the expression of TNFα-mRNA in peritoneal macrophages after application of extracts such as walnut peels. METHODS: Male and female mice were randomly divided into groups and macrophage smears were prepared after oral administration; PGEM-1 plasmids containing hTNF cDNA were transformed into EcoliJM 103, and positive clones were selected for cell culture and amplification; and digoxigenin-labeled probes were prepared by random primer method. The expression of TNFα-mR NA in macrophages was detected by in situ hybridization. RESULTS: Anti-tumor extracts such as walnut peels could enhance the expression of TNFα-mRNA in mouse peritoneal macrophages. Conclusion: This project has revealed the anti-tumor mechanism of anti-tumor herbs such as walnut peel and other extracts.