Arbustini E, Diegoli M, Fasani R, Grasso M, Morbini P, Banchieri N, Bellini O, Dal Bello B, Pilotto A, Magrini G, Campana C, Fortina P, Gavazzi A, Narula J, Viganò M
Cardiovascular Pathology and Molecular Diagnostic, Istituto di Ricovero e Cura a Carattere Scientifico Policlinico San Matteo, Pavia, Italy.
Am J Pathol. 1998 Nov;153(5):1501-10. doi: 10.1016/S0002-9440(10)65738-0.
Mitochondrial (mt)DNA defects, both deletions and tRNA point mutations, have been associated with cardiomyopathies. The aim of the study was to determine the prevalence of pathological mtDNA mutations and to assess associated defects of mitochondrial enzyme activity in dilated cardiomyopathy (DCM) patients with ultrastructural abnormalities of cardiac mitochondria. In a large cohort of 601 DCM patients we performed conventional light and electron microscopy on endomyocardial biopsy samples. Cases with giant organelles, angulated, tubular, and concentric cristae, and crystalloid or osmiophilic inclusion bodies were selected for mtDNA analysis. Mutation screening techniques, automated DNA sequencing, restriction enzyme digestion, and densitometric assays were performed to identify mtDNA mutations, assess heteroplasmy, and quantify the amount of mutant in myocardial and blood DNA. Of 601 patients (16 to 63 years; mean, 43.5 +/- 12.7 years), 85 had ultrastructural evidence of giant organelles, with abnormal cristae and inclusion bodies; 19 of 85 (22.35%) had heteroplasmic mtDNA mutations (9 tRNA, 5 rRNA, and 4 missense, one in two patients) that were not found in 111 normal controls and in 32 DCM patients without the above ultrastructural mitochondrial abnormalities. In all cases, the amount of mutant was higher in heart than in blood. In hearts of patients that later underwent transplantation, cytochrome c oxidase (Cox) activity was significantly lower in cases with mutations than in those without or controls (P = 0.0008). NADH dehydrogenase activity was only slightly reduced in cases with mutations (P = 0.0388), whereas succinic dehydrogenase activity did not significantly differ between DCM patients with mtDNA mutations and those without or controls. The present study represents the first attempt to detect a morphological, easily identifiable marker to guide mtDNA mutation screening. Pathological mtDNA mutations are associated with ultrastructurally abnormal mitochondria, and reduced Cox activity in a small subgroup of non-otherwise-defined, idiopathic DCMs, in which mtDNA defects may constitute the basis for, or contribute to, the development of congestive heart failure.
线粒体(mt)DNA缺陷,包括缺失和tRNA点突变,都与心肌病有关。本研究的目的是确定病理性mtDNA突变的患病率,并评估心脏线粒体超微结构异常的扩张型心肌病(DCM)患者线粒体酶活性的相关缺陷。在601例DCM患者的大样本队列中,我们对心内膜活检样本进行了传统的光学和电子显微镜检查。选择具有巨大细胞器、角状、管状和同心嵴以及晶体或嗜锇包涵体的病例进行mtDNA分析。采用突变筛查技术、自动DNA测序、限制性酶切和密度测定法来鉴定mtDNA突变、评估异质性,并定量心肌和血液DNA中突变体的数量。601例患者(年龄16至63岁;平均43.5±12.7岁)中,85例有巨大细胞器的超微结构证据,伴有异常嵴和包涵体;85例中的19例(22.35%)有异质性mtDNA突变(9个tRNA、5个rRNA和4个错义突变,两名患者中有一人出现),在111名正常对照者和32例无上述线粒体超微结构异常的DCM患者中未发现这些突变。在所有病例中,心脏中的突变体量高于血液中的。在后来接受移植的患者心脏中,有突变的病例中细胞色素c氧化酶(Cox)活性显著低于无突变病例或对照者(P = 0.0008)。有突变的病例中NADH脱氢酶活性仅略有降低(P = 0.0388),而mtDNA突变的DCM患者与无突变患者或对照者之间琥珀酸脱氢酶活性无显著差异。本研究首次尝试检测一种形态学上易于识别的标志物,以指导mtDNA突变筛查。病理性mtDNA突变与超微结构异常的线粒体有关,并且在一小部分未明确病因的特发性DCM中Cox活性降低,其中mtDNA缺陷可能构成充血性心力衰竭发生的基础或促成因素。