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对接受膝关节和髋关节置换术的老年患者皮质骨中线粒体DNA的Southern分析。

Southern analysis of mitochondrial DNA in cortical bone of elderly patients undergoing knee and hip arthroplasty.

作者信息

Varanasi S S, Datta H K

机构信息

School of Clinical and Laboratory Sciences, Department of Clinical Biochemistry, The Medical School, University of Newcastle upon Tyne, Newcastle upon Tyne, NE2 4HH, UK.

出版信息

J Pathol. 2001 Apr;193(4):557-62. doi: 10.1002/path.823.

Abstract

The role of mitochondrial DNA deletions (dmtDNA) in involutional bone loss seen in elderly men and women has never been examined. The present investigation was carried out to determine the extent of dmtDNA in cortical bone of elderly patients undergoing knee and hip arthroplasties. The majority of earlier studies have employed the polymerase chain reaction (PCR) to detect and quantify dmtDNA in different body tissues. In the present study, Southern blotting was used to screen bone biopsies from 30 patients undergoing orthopaedic surgery (mean age+/-SD 67.5+/-9.6 years; range 49-87 years). The blotting of PvuII-digested genomic DNA, carried out using mtDNA probes covering the entire span of mtDNA, revealed high levels of deletions in six subjects (mean age+/-SD 63.0+/-10.1 years; range 49-78 years) and moderate to low levels of mutations in another 14 subjects (mean age+/-SD 64.9+/-8.9 years; range 53-87 years). The importance of this rather high prevalence of dmtDNA in the bone of the elderly is discussed in terms of possible involvement of increased production of oxygen-derived free radicals and oxidative stress, and its possible role in the accelerated bone loss leading to osteoporosis.

摘要

线粒体DNA缺失(dmtDNA)在老年男性和女性出现的退行性骨质流失中的作用从未被研究过。本研究旨在确定接受膝关节和髋关节置换术的老年患者皮质骨中dmtDNA的程度。大多数早期研究采用聚合酶链反应(PCR)来检测和量化不同身体组织中的dmtDNA。在本研究中,Southern印迹法用于筛查30例接受骨科手术患者(平均年龄±标准差67.5±9.6岁;范围49 - 87岁)的骨活检样本。使用覆盖整个线粒体DNA跨度的线粒体DNA探针,对经PvuII消化的基因组DNA进行印迹分析,结果显示6名受试者(平均年龄±标准差63.0±10.1岁;范围49 - 78岁)存在高水平的缺失,另外14名受试者(平均年龄±标准差64.9±8.9岁;范围53 - 87岁)存在中度至低度的突变。从氧衍生自由基生成增加和氧化应激的可能参与,以及其在导致骨质疏松症的加速骨质流失中的可能作用方面,讨论了dmtDNA在老年人骨骼中这种相当高的患病率的重要性。

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