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大鼠脑和股四头肌组织中mRNA表达及组织形态的死后变化

Postmortem Changes in mRNA Expression and Tissue Morphology in Brain and Femoral Muscle Tissues of Rat.

作者信息

Choi Sujin, Jung Minju, Jeong Mingyoung, Kim Sohyeong, Lee Dong Geon, Park Kwangmin, Xuan Xianglan, Park Heechul, Kim Dong Hyeok, Kim Jungho, Lee Min Ho, Cho Yoonjung, Kim Sunghyun

机构信息

Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea.

Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea.

出版信息

Int J Mol Sci. 2025 Jul 22;26(15):7059. doi: 10.3390/ijms26157059.

Abstract

The postmortem interval (PMI), defined as the time elapsed between death and the discovery or examination of the body, is a crucial parameter in forensic science for estimating the time of death. There are many ways to measure the PMI, such as Henssge's nomogram, which uses rectal temperature measurement; livor mortis; rigor mortis; and forensic entomology. However, these methods are usually affected by various conditions in the surrounding environment. The purpose of the present study was to compare molecular genetics and histological changes in the brain and skeletal muscle tissues of SD rats over increasing periods of time after death. For the PMIs, we considered 0 h, 6 h, 12 h, 24 h, 36 h, 48 h, 4 days, 6 days, 8 days, 10 days, 14 days, and 21 days and compared them at 4 °C and 26 °C. Hematoxylin and Eosin (H&E) staining was performed to observe tissue changes. Morphological tissue changes were observed in cells for up to 21 days at 4 °C, and cell destruction was visually confirmed after 14 days at 26 °C. Total RNA (tRNA) was isolated from each tissue sample, and complementary DNA (cDNA) was synthesized. A reverse transcription quantitative PCR (RT-qPCR) SYBR Green assay targeting three types of housekeeping genes, including , , , and 5S rRNA, was performed. The results showed that and 5S rRNA were highly stable and could be better RNA targets for estimating the PMI in brain and skeletal muscle tissues. Conversely, and showed poor stability and low expression levels. In conclusion, these molecular biomarkers could be used as auxiliary indicators of the PMI in human, depending on the stability of the marker.

摘要

死后间隔时间(PMI)定义为死亡与尸体发现或检查之间经过的时间,是法医学中估计死亡时间的关键参数。测量PMI有多种方法,如亨斯格诺谟图,它利用直肠温度测量;尸斑;尸僵;以及法医昆虫学。然而,这些方法通常会受到周围环境各种条件的影响。本研究的目的是比较SD大鼠死后不同时间段大脑和骨骼肌组织中的分子遗传学和组织学变化。对于PMI,我们考虑了0小时、6小时、12小时、24小时、36小时、48小时、4天、6天、8天、10天、14天和21天,并在4℃和26℃下进行比较。进行苏木精和伊红(H&E)染色以观察组织变化。在4℃下,细胞形态学组织变化观察长达21天,在26℃下,14天后肉眼确认细胞破坏。从每个组织样本中分离总RNA(tRNA),并合成互补DNA(cDNA)。进行了针对三种管家基因(包括 、 、 和5S rRNA)的逆转录定量PCR(RT-qPCR)SYBR Green分析。结果表明, 和5S rRNA高度稳定,可作为估计大脑和骨骼肌组织中PMI的更好的RNA靶标。相反, 和 稳定性差且表达水平低。总之,根据标志物的稳定性,这些分子生物标志物可作为人类PMI的辅助指标。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b19e/12346364/6073e77e3989/ijms-26-07059-g001.jpg

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