摘要
目的应用实时定量PCR技术检测大鼠皮肤挫伤后NF-κB mRNA时序性表达规律,分析其与损伤时间的关系。方法制备大鼠皮肤挫伤模型,于伤后0.5,1,6,12,18,24,30h取材,一步法提取总RNA,检测其完整性和浓度,按照0.4μg总RNA量逆转录合成第一链cDNA,以rlp32为内参基因进行荧光定量检测,采用△C t法比较其与正常皮肤组织NF-κB mRNA表达的倍数关系。结果皮肤挫伤组织中NF-κB mRNA伤后0.5h表达显著增强(P<0.05),6h出现高峰,12h降至正常水平,随后略有回升。结论大鼠皮肤挫伤组织NF-κB mRNA的表达,随着损伤经过时间的延长呈规律性变化。
Objective To investigate the expression of NF-κB mRNA during contused skin in rats and to explore the relationship between NF-κB mRNA with the wound age. Methods Forty-eight rats were divided into two groups randomly： contusion group （7 subgroups） and control group, the contusions was performed on the right posterior limb of rat, the samples were extracted respectively at 0. 5 ,1,6,12,18 ,24 and 30h after contusion. Total RNA were isolated from skin of two groups and reverse transcription polymerase chain reaction was conducted to synthesize the 1 -st strand cDNA. The initial amount of NF-κB mRNA in each sample was evaluated using the Real-time PCR by means of cDNA as template. Results After contusion there was a rapid increase in NF-κB mRNA which peaked at 6. Oh in skin. Conclusion This study demonstrated the possible use of NF-κB mRNA analysis for forensic examination of wound age because of its time-order expression after contusion in skin.
出处
《中国法医学杂志》
CSCD
2008年第6期370-373,共4页
Chinese Journal of Forensic Medicine
基金
山西省留学回国人员基金资助项目(200528)
山西省青年科技研究基金项目(2007021047)
山西省教育厅高校科技开发项目(200713007)
