大鼠脑、肝组织3种RNA相对表达量与死亡时间的关系

发布时间：2019-06-28 17:19

【摘要】：目的研究大鼠脑、肝脏组织3种RNA相对表达量与死亡时间的关系,比较两组织用于死亡时间推断的适用性。方法将48只Wistar大鼠随机分为6组,断颈处死,分别于死后0、24、48、72、120、168 h取大鼠脑皮质及肝右外叶。提取脑、肝组织总RNA,采用实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-qPCR)技术检测两组织中miR(脑特异性miR-124、肝特异性miR-122)及甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase,GAPDH)、β-肌动蛋白(β-actin)mRNA的循环阈值(cyclic threshold,Ct)。用Best Keeper程序筛选内参,对目的RNA的Ct值进行2~(-ΔΔC)t转换,用SPSS及Graph Pad软件对其进行单因素方差分析,并对ΔCt值进行曲线回归分析。结果 miR-124和miR-122分别为脑、肝组织的内参。脑组织GAPDH、β-actin mRNA的相对表达量在死后72 h内逐渐增加,并于死后72 h达到峰值,之后逐渐下降(P0.05);而肝GAPDH、β-actin mRNA的相对表达量从死后48 h开始逐渐下降(P0.05)。所有分子的最适回归模型均为三次模型,且肝组织的相关系数r大于脑组织。结论大鼠脑、肝组织中GAPDH、β-actin mRNA的相对表达量与死亡时间具有相关性,肝组织比脑组织更适用于死亡时间推断。
[Abstract]:Objective to study the relationship between the relative expression of three kinds of RNA in rat brain and liver and the time of death, and to compare the applicability of the two tissues in predicting the time of death. Methods 48 Wistar rats were randomly divided into 6 groups. The cerebral cortex and right lateral lobe of liver were taken at 0 h, 24 h, 48 h and 721 20168 h after death. Total RNA, was extracted from brain and liver tissue by real-time fluorescence quantitative PCR (real-time fluorescence quantitative PCR,RT-qPCR) to detect miR (brain-specific miR-124, liver-specific miR-122), glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase,GAPDH) and 尾-actin (尾-actin) mRNA) circulation threshold (cyclic threshold,Ct) in the two tissues. The internal parameters were screened by Best Keeper program, the KT value of the target RNA was transformed by 2 ~ (- 螖 C) t), the one-factor variance analysis was carried out by SPSS and Graph Pad software, and the curve regression analysis of 螖 C _ t value was carried out. Results miR-124 and miR-122 were internal parameters of brain and liver, respectively. The relative expression of GAPDH, 尾-actin mRNA in brain tissue increased gradually within 72 hours after death, reached the peak at 72 hours after death, and then decreased gradually (P 0.05), while the relative expression of GAPDH, 尾-actin mRNA in liver began to decrease gradually from 48 hours after death (P 0.05). The optimal regression model of all molecules was cubic model, and the correlation coefficient of liver tissue was higher than that of brain tissue. Conclusion the relative expression of GAPDH, 尾-actin mRNA in rat brain and liver tissue is correlated with the time of death, and the relative expression of 尾-actin mRNA in liver tissue is more suitable for the inference of death time than in brain tissue.
【作者单位】：青海大学医学院;青海省公安厅刑事技术研究管理中心;
【分类号】：D919

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