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目的：研究槟榔碱(arecoline,ARC)体内对小鼠骨髓细胞DNA的损伤作用，并探讨其作用机制。方法：采用单细胞凝胶电泳实验分析ARC对小鼠骨髓细胞DNA损伤作用。采用流式细胞术、DCFH-DA荧光染色、罗丹明123染色等方法对小鼠骨髓细胞的细胞周期，超氧化物歧化酶(superoxide dismutase,SOD)活性和谷胱甘肽(glutathione,GSH)、丙二醛(malondial dehyde,MDA)、活性氧(reactive oxygen species,ROS)含量，线粒体膜电位(Δψm),p53蛋白含量进行分析。结果：给予ARC1/2 LD50剂量可引起小鼠骨髓细胞DNA损伤，与空白组相比拖尾现象较严重。随着ARC给药剂量的增加，G0/G1期的细胞比例明显增加(P<0.01)，使骨髓细胞周期停滞在G0/G1期。ARC作用小鼠骨髓细胞后，SOD活性和GSH含量明显降低(P<0.01),MDA和ROS含量明显升高(P<0.01)，并且随着ARC浓度增加，Δψm明显降低(P<0.01)。ARC给药组骨髓细胞内p53蛋白含量随给药剂量的增加而明显升高(P<0.01)，呈一定的剂量依赖关系。结论：ARC能造成小鼠骨髓细胞DNA损伤，可增加p53蛋白表达诱导小鼠骨髓细胞周期阻滞在G0/G1期，引起Δψm降低，ROS增加，脂质过氧化增强，抗氧化功能受到严重损害，氧化和抗氧化失衡，这些是ARC引起小鼠骨髓细胞DNA损伤的机制。
Abstract：Objective: To study the DNA damage effect of arecoline(ARC) on mouse bone marrow cells in vivo, and to explore its mechanism. Methods: Single cell gel electrophoresis was used to analyze the DNA damage effect of ARC on mouse bone marrow cells. Flow cytometry, DCFH-DA fluorescence staining, and Rhodamine 123 staining were used to analyze the cell cycle, superoxide dismutase(SOD) activity,glutathione(GSH),malondial dehyde(MDA),and reactive oxygen species(ROS) content, mitochondrial transmembrane potential (Δψm) and p53 protein content of mouse bone marrow cells. Results: ARC, at a dose of 1/2 LD50, could induce DNA damage in mouse bone marrow cells. The difference was significant compared to the negative control group. Cell cycle changed observably in a dose-dependent manner when exposed to ARC. The percentage of cells in G0/G1 phase was significantly increased(P<0.01), and ARC caused G0/G1 phases′ cell cycle arrest in mouse bone marrow cells. After the administration of ARC, SOD activity and GSH content were significantly decreased(P<0.01), MDA and ROS contents were significantly increased(P<0.01). Furthermore, the Δψm was significantly reduced with increasing dose(P<0.01). The protein levels of p53 in mouse bone marrow cells was increased in a dose-dependent manner after ARC treatment(P<0.01). Conclusion: ARC can cause DNA damage of mouse bone marrow cells, increase the expression of p53 protein causing cell cycle arrest in the G0/G1 phase, decrease Δψm, increase ROS in mouse bone marrow cells, enhance lipid peroxidation, severely damage cellular antioxidant function,and lead to the imbalance of oxidation and antioxidation. These are the mechanisms of DNA damage in mouse bone marrow cells caused by ARC.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20250225.001
中图分类号:R285.5
引用信息:
[1]于蕾,于晶涵,宋辉,等.基于周期阻滞和氧化-抗氧化失衡研究槟榔碱致小鼠骨髓细胞DNA损伤的机制[J].海南医科大学学报,2026,32(02):89-95.DOI:10.13210/j.cnki.jhmu.20250225.001.
基金信息:
国家自然科学基金（82060778）; 海南省自然科学基金（820RC776）; 海南省卫生健康科技创新联合项目（WSJK2024MS162）; 黑龙江省中医药科研课题项目（ZHY2024-098）~~
2025-02-25
2025-02-25
2025-02-25