彗星试验检测大鼠死后肾脏细胞DNA含量与死亡时间的关系  

Relationship between DNA of Rat Kidney and Post by Comet Assay

作　　者：靳俊峰[1] 高翠莲[2] 陈玉川[2] 罗光华[2] 

机构地区：[1]遵义医学院珠海校区病理教研室,广东珠海519041 [2]中山大学中山医学院法医学系,广东广州510080

出　　处：《中山大学学报（医学科学版）》2009年第4期386-389,共4页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金(036628)

摘　　要：【目的】探讨特定条件下,利用彗星试验测定SD大鼠死后6～48h肾脏细胞DNA含量与死亡时间的关系。【方法】健康成年雌性SD大鼠,按取材时间随机分为6组,每组3只。颈椎脱臼法处死动物,置于25.1℃(广州地区近5年10月份平均气温)培养箱中,分别于0、6、12、24、36、48、60h取大鼠肾脏组织,制备单细胞悬液,进行彗星试验,荧光显微成像系统采集图像,采集相关参数(Comet4.0)并进行Kruskal-Wallis检验。【结果】死后6～48h,SD大鼠肾脏细胞DNA碎片随死亡时间延长而增加,尾长在死后各时间点依次呈明显的增长趋势,Oliver尾矩和尾DNA随死亡时间的延长,亦有一定的增长趋势,60h未能测出。Kruskal-Wallis检验显示:各时间点彗星尾长均数的差异有高度显著性(P<0.01);彗星尾矩均数在死后6h和12h差异无统计学意义(P>0.05),24h和36h彗星尾矩均数差异有显著性(P<0.05);尾DNA均数在6h,12h,24h三个时间点差异无显著性(P>0.05),但与36h,48h尾DNA均数的差异均有显著性(P<0.05)。【结论】SD大鼠肾脏细胞DNA降解随死亡时间延长而增加,彗星试验技术为死亡时间推断提供了重要的实验依据。[Objective] To study the correlation between kidney cell DNA degradation and postmortem interval within the span of 6-48 hour after the subject rats＇ death. [Methods] To select 18 healthy mature female SD rats and equally divide them into 6 groups. To execute the rats with cervical spine articulation and put the rats under the incubator temperature of 25.1℃ （the average temperature of the 5 previous Decembers in Guangzhou prefecture）. Sample kidney tissue from the rat separately 0 hour, 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, and 60 hours after the rats＇ execution to prepare monoplast suspension, which is committed to comet assay. The comet images were captured by fluorescence CCD. Kinetic Comet 4.0 software was used to analyze images. Relevant data were collected by kinetic Comet 4.0 software and were subjected to Kruskal-Wallis test. [Results] Within the postmortem interval of 6-48 h, the number of SD rat kidney cell DNA fragments increased as the postmortem interval lengthens. So did the comet tail length. The Oliver tail moment and tail DNA of comet also showed sign of increase in positive proportion to the postmortem interval （their values corresponding to 60-hour-postmortem-interval were not obtainable. Kruskal-Wallis test indicated： the discrepancies of TL among the 6 groups were all significant （P 〈 0.01）. The difference of TM between 6 h group and 12 h group was not significant （P 〉 0.05）. The difference of TM between 24 h and 36 h was significant （P 〈 0.05）. The difference of TDNA among 6 h, 12 h, and 36 h groups were not significant （P 〉 0.05）. The difference of TDNA between 36 h and 48 h was significant （P 〈 0.05）. [Conclusion] Degradation of nuclear DNA of the rat kidney cells increases as the postmortem interval lengthens and comet assay may provide important empirical evidence for determining the postmortem interval.

关 键 词：DNA降解 彗星试验 死亡时间 肾脏细胞 

分 类 号：R64[医药卫生—外科学] R446.1[医药卫生—临床医学]