摘要
目的:通过检测活性氮族引起的核酸损伤标志物8-硝基鸟嘌呤在砷暴露小鼠肾脏组织中的表达情况,分析三氧化二砷对肾组织RNA损伤的作用途径。方法:实验于2005-11/2006-01在大连医科大学毒理学实验室完成。选用昆明种健康小鼠40只,按体质量随机分为4组,即正常对照组、5μmol/L、10μmol/L和20μmol/L三氧化二砷染毒组,每组10只。各染砷组小鼠通过饮用含不同剂量三氧化二砷自来水的方式使小鼠暴露砷,连续染砷60d。正常对照组每天摄入相同剂量的生理盐水。染砷60d后处死各组小鼠,立即取肾脏皮质,应用苏木精-伊红染色法进行肾皮质组织病理学观察;应用免疫组织化学及图像分析的方法检测肾组织内8-硝基鸟嘌呤的表达,根据8-硝基鸟嘌呤表达的吸光度值进行定量分析。结果:40只小鼠全部进入结果分析,无脱失。①各染砷组小鼠肾脏皮质区肾小管细胞出现肿胀,胞质内有空泡变性,核固缩、溶解等明显的病理学变化;而正常对照组小鼠的肾脏细胞未见上述病理变化。②5μmol/L、10μmol/L和20μmol/L三氧化二砷染毒组小鼠肾组织细胞的抗8-硝基鸟嘌呤免疫染色平均吸光度显著大于正常对照组(0.027±0.014,0.039±0.015,0.043±0.014,0.004±0.002,P<0.05),20μmol/L三氧化二砷染毒组与5μmol/L三氧化二砷染毒组相比,吸光度明显增加(P<0.05)。结论:活性氮族可能参与了砷对肾脏细胞核酸的损伤作用,8-硝基鸟嘌呤可能是砷致肾损伤的标志物。
AIM: To address the molecular mechanism of DNA damage by arsenic trioxide (As2O3) in kidney by detecting the expression of 8-nitroguanine (8-NOrG), which is the biochemical marker of nitrative nuclear acid damage.
METHODS: The experiments were carried out in the Laboratory of Toxicology, Dalian Medical University between November 2005 and January 2006. Forty Kunming species mice were divided into 4 groups, 10 rats in each, including a control group and three As203 groups. Different concentrations of As203 (5, 10, 20 μmol/L) were administered to mice for consecutive 60 days, followed by cervical dislocation. Physiological saline was given in the control group. Kidney cortex was sampled to detect the pathologic changes following hematoxylin-eosin staining; Immunohistochemical methods and image analysis were used to investigate 8-NOrG expression in kidney of mice, followed by the quantitative analysis Of the absorbance of slices.
RESULTS: All 40 mice, which had entered the study, were involved in the evaluation.(1)Pathological changes were found in the 5 μmol/L, 10 μmol/L and 20 μmol/L of As203 groups. The nephric tubule cells in the cortical area of the mice had become swollen, vacuolar degeneration occurred in the cytoplasm, additional pathological changes such as karyopycnosis and caryolysis also occurred. (2)The absorbance of control group was significantly lower than those of the As203 treated groups (0.027±0.014, 0.039±0.015, 0.043±0.014, 0.004±0.002, P 〈 0.05). Compared to the 5 mol/L As203 group, the expression of 8-NOrG was increased significantly in the 20 μmol/L As2O3 group (P 〈 0.05).
CONCLUSION: Reactive nitrogen species may contribute to DNA damage in the kidney. And 8-NOrG may be the early marker of kidney damage induced by arsenic trioxide.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第25期4906-4908,共3页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家自然科学基金(30571584
30600488)
辽宁省教育厅项目(05L113)~~