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脑内 MMPs、CLDN5、OCLN、ZO1 和 AQP 的表达增加与致命性热射病后脑水肿有关。

Increased cerebral expressions of MMPs, CLDN5, OCLN, ZO1 and AQPs are associated with brain edema following fatal heat stroke.

机构信息

Department of Forensic Medicine, National Police University of China, Shenyang, China.

Department of Forensic Pathology, School of Forensic Medicine, Southern Medical University, Guangzhou, China.

出版信息

Sci Rep. 2017 May 10;7(1):1691. doi: 10.1038/s41598-017-01923-w.

DOI:10.1038/s41598-017-01923-w
PMID:28490769
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5431794/
Abstract

Human brain samples were collected from 46 autopsy cases, including 23 fatal heat stroke cases and 23 age-matched controls. Nine candidate reference genes (PES1, POLR2A, IPO8, HMBS, SDHA, GAPDH, UBC, B2M, ACTB) were evaluated in the cerebral cortex of 10 forensic autopsy cases (5 heat stroke and 5 controls), using the geNorm module in qBase software. SDHA, POLR2A, IPO8 and HMBS were identified as the most stable reference genes. Using these validated reference genes, mRNA expressions of Matrix metalloproteinases (MMPs, MMP2 and MMP9), Claudin5 (CLDN5), Occludin (OCLN), Zona occludens protein-1 (ZO1) and Aquaporins (AQPs, AQP1 and AQP4) in the cerebral cortex were examined. Relative mRNA quantification using Taqman real-time PCR assay demonstrated increased calibrated normalized relative quantity (CNRQ) values of MMP9, CLDN5, OCLN, ZO1 and AQP4 in heat stroke cases. Heat stroke cases showed an increase in brain water content, which was found to be positively correlated with MMP9, OCLN, ZO1 and CLDN5 mRNA. When using one conventional reference gene (GAPDH or ACTB) for normalization, no difference was detected between heat stroke and controls. In immunostaining, only AQP4 showed more intense staining in most heat stroke cases. The present study, for the first time, reports increased cerebral MMP9, CLDN5, OCLN, ZO1 and AQP4 in heat stroke and suggest a crucial role of reference gene selection when using postmortem human tissues.

摘要

从 46 例尸检病例中收集了人脑样本，包括 23 例致命性热射病病例和 23 例年龄匹配的对照病例。在 10 例法医尸检病例（5 例热射病和 5 例对照）的大脑皮层中，使用 qBase 软件中的 geNorm 模块评估了 9 个候选参考基因（PES1、POLR2A、IPO8、HMBS、SDHA、GAPDH、UBC、B2M、ACTB）。SDHA、POLR2A、IPO8 和 HMBS 被确定为最稳定的参考基因。使用这些经过验证的参考基因，检测了大脑皮层中基质金属蛋白酶（MMPs、MMP2 和 MMP9）、Claudin5（CLDN5）、Occludin（OCLN）、紧密连接蛋白-1（ZO1）和水通道蛋白（AQPs、AQP1 和 AQP4）的 mRNA 表达。Taqman 实时 PCR 检测的相对 mRNA 定量显示，热射病病例的 MMP9、CLDN5、OCLN、ZO1 和 AQP4 的校准标准化相对数量（CNRQ）值增加。热射病病例的脑水含量增加，与 MMP9、OCLN、ZO1 和 CLDN5 mRNA 呈正相关。当使用一个常规参考基因（GAPDH 或 ACTB）进行归一化时，热射病病例和对照组之间没有差异。在免疫染色中，只有 AQP4 在大多数热射病病例中显示出更强的染色。本研究首次报道了热射病中大脑 MMP9、CLDN5、OCLN、ZO1 和 AQP4 的增加，并表明在使用死后人体组织时，参考基因选择至关重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/66d880aacba5/41598_2017_1923_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/158629c94069/41598_2017_1923_Fig1_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/748a16299872/41598_2017_1923_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/d1044e0b2c9b/41598_2017_1923_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/66d880aacba5/41598_2017_1923_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/158629c94069/41598_2017_1923_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/337aff4c01cc/41598_2017_1923_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/6e6535decdb3/41598_2017_1923_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/4924a1998bbf/41598_2017_1923_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/748a16299872/41598_2017_1923_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/d1044e0b2c9b/41598_2017_1923_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e8d4/5431794/66d880aacba5/41598_2017_1923_Fig7_HTML.jpg

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