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目的探讨小白鼠死后脾组织GAPDH mRNA和β-actin mRNA降解情况与死亡时间(PMI)的关系。方法 48只NIH小白鼠安乐死处死,分别置于10℃和25℃温控系统内,利用两步法RT-PCR技术和核酸蛋白测定仪定量cDNA方法检测小白鼠脾GAPDH mRNA和β-actin mRNA在死后即刻至72h降解情况。结果在10℃温控系统内的小白鼠死后即刻至72h脾组织均可检测到GAPDH mRNA和β-actin mRNA,且其扩增产物呈规律性下降趋势。25℃温控系统内的小白鼠死后即刻至48h脾组织均可检测到GAPDH mRNA和β-actin mRNA,且其扩增产物呈规律性下降趋势。结论小白鼠死亡后脾GAPDH mRNA和β-actin mRNA降解与PMI负相关,可为PMI推断提供一种新的观测指标。
Objective To investigate the relationship between the degradation of GAPDH mRNA and β-actin mRNA and the time to death (PMI) in mice spleen after death. Methods Forty-eight NIH mice were euthanized and placed in 10 ℃ and 25 ℃ temperature control systems respectively. The expression of GAPDH mRNA and β-actin mRNA in spleen of mice were detected by two-step RT-PCR and nucleic acid protein analyzer Immediately after death to 72h degradation. Results The GAPDH mRNA and β-actin mRNA were detected in the spleen tissue of mice at 10 ℃ immediately after death, and the amplification products showed a regular decreasing trend. The GAPDH mRNA and β-actin mRNA were detected in the spleen of mice at 25 ℃ immediately after death, and the amplification products showed a regular decreasing trend. Conclusion The expression of GAPDH mRNA and β-actin mRNA in the spleen of mice is negatively correlated with PMI, which may provide a new observation index for PMI inference.