大鼠溺死内参基因筛选和水通道蛋白5的表达

Reference genes selection and expression of water channel protein 5 for drowning rats

摘要:

目的：在溺死尸体不同时间点模型中，筛选大鼠稳定表达的内参基因和探讨水通道蛋白5 mRNA（AQP-5 mRNA）的表达。方法：先建立溺死尸体不同时间点模型（0 min，n=6；15 min，n=6；4h，n=6；16h，n=6；48 h，n=6；72 h，n=6），提取样本总的RNA,逆转录合成cDNA，qRT-PCR定量候选内参基因，geNorm和NormFinder排序各候选内参基因的表达，筛选出最稳定的内参基因，最后定量目的基因AQP-5 mRNA的表达。结果：该模型中筛选出ACTB为最稳定的内参基因，除72 h时间点，AQP-5 mRNA的表达均显著下调（15 min/0 min=0.51，P<0.05；4 h/0 min=0.53，P<0.05；16 h/0 min=0.63，P<0.05；48 h/0 min=0.81，P<0.05；72 h/0 min=0.89，P >0.05）。结论：本研究AQP-5 mRNA在大鼠溺死表达有一定阳性意义，有助于法医溺死鉴定.

Abstract:

Objective：To select stable expression of reference gene and to explore the water channel protein 5 mRNA expression for drowning rat model in different time points. Methods：Drowning rat was modeled in different time points（0 min，n=6；15 min，n=6；4 h，n=6；16 h，n=6；48 h，n=6；72 h，n=6）. Total RNA was extracted from samples and reversed transcription was conducted for cDNA syn－thesis. Candidate reference genes were detected by real-time PCR. In order to screen out the most stable reference gene，geNorm and NormFinder was used to rank the expression of each gene. Then，AQP-5 mRNA expression was detected by qRT-PCR. Results：ACTB was the most stable reference gene. Our study showed that the expression of AQP-5 mRNA was downloaded except 72 h in this model（15 min/0 min=0.51，P<0.001；4 h/0 min=0.53，P<0.001；16 h/0 min=0.63，P<0.001；48 h/0 min=0.81，P<0.05；72 h/0 min=0.89，P>0.05）. Conclusion：AQP-5 mRNA expression has some positive meaning and improves forensic identification in drowning model.