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目的研究SD大鼠脑、心肌和肾组织细胞内β-actin mRNA的降解与早期死亡时间的关系,为早期死亡时间的推断寻找新的指标。方法大鼠处死后置于20℃的环境中,分别于死后不同时间点提取脑、心肌、肾的总RNA,采用实时荧光定量RT-PCR方法检测总RNA中β-actin mRNA的水平(Ct值),分析死后经过时间与Ct值的线性关系,并建立推断早期死亡时间的线性回归方程。结果随死亡时间的延长,3种组织内β-actin mRNA的水平均发生了显著的下降,心肌和肾组织中β-actin mRNA死后24h内的降解速率更显著,而脑组织中β-actin mRNA的降解速率变化不明显。各组织的Ct值变化与死亡时间之间存在一定的线性关系。结论大鼠死亡早期脑、心肌和肾组织内β-actin mRNA降解明显,可以通过其降解规律来推断早期死亡时间。
Objective To study the relationship between the degradation of β-actin mRNA and the time of early death in the brain, myocardium and kidney of SD rats and to find a new index for the estimation of the early death time. Methods After sacrificed, the rats were placed in the environment at 20 ℃. The total RNA of brain, myocardium and kidney were extracted at different time points after death. The level of β-actin mRNA in total RNA was detected by real-time fluorescence quantitative RT-PCR Value), the linear relationship between the time elapsed after death and the Ct value was analyzed, and a linear regression equation was established to estimate the early death time. Results With the prolongation of death time, the levels of β-actin mRNA in three kinds of tissues all decreased significantly. The degradation rate of β-actin mRNA in myocardium and kidney was more significant within 24h after death, The rate of mRNA degradation did not change significantly. There was a certain linear relationship between the change of Ct value and death time in each organization. Conclusion The β-actin mRNA in brain, myocardium and kidney of rats is obviously degraded in early stage of death, and the early death time can be deduced through its degradation.