外周动脉疾病患者骨骼肌中线粒体呼吸异常。

Abnormal mitochondrial respiration in skeletal muscle in patients with peripheral arterial disease.

作者信息

Pipinos Iraklis I, Sharov Victor G, Shepard Alexander D, Anagnostopoulos Petros V, Katsamouris Asterios, Todor Anastasia, Filis Konstantinos A, Sabbah Hani N

机构信息

Department of Surgery, University of Nebraska Medical Center, 983280 Nebraska Medical Center, Omaha, NE 68198-3280, USA.

出版信息

J Vasc Surg. 2003 Oct;38(4):827-32. doi: 10.1016/s0741-5214(03)00602-5.

DOI:10.1016/s0741-5214(03)00602-5

PMID:14560237

Abstract

OBJECTIVE

Discrete morphologic, enzymatic and functional changes in skeletal muscle mitochondria have been demonstrated in patients with peripheral arterial disease (PAD). We examined mitochondrial respiration in the gastrocnemius muscle of nine patients (10 legs) with advanced PAD and in nine control patients (nine legs) without evidence of PAD.

METHODS

Mitochondrial respiratory rates were determined with a Clark electrode in an oxygraph cell containing saponin-skinned muscle bundles. Muscle samples were obtained from the anteromedial aspect of the gastrocnemius muscle, at a level 10 cm distal to the tibial tuberosity. Mitochondria respiratory rate, calculated as nanoatoms of oxygen consumed per minute per milligram of noncollagen protein, were measured at baseline (V(0)), after addition of substrates (malate and glutamate; (V(SUB)), after addition of adenosine diphosphate (ADP) (V(ADP)), and finally, after adenine nucleotide translocase inhibition with atractyloside (V(AT)). The acceptor control ratio, a sensitive indicator of overall mitochondrial function, was calculated as the ratio of the respiratory rate after the addition of ADP to the respiratory rate after adenine nucleotide translocase inhibition with atractyloside (V(ADP)/ V(AT)).

RESULTS

Respiratory rate in muscle mitochondria from patients with PAD were not significantly different from control values at baseline (0.31 +/- 0.06 vs 0.55 +/- 0.12; P =.09), but V(sub) was significantly lower in patients with PAD compared with control subjects (0.43 +/- 0.07 vs 0.89 +/- 0.20; P <.05), as was V(ADP) (0.69 +/- 0.13 vs 1.24 +/- 0.20; P <.05). Respiratory rates after atractyloside inhibition in patients with PAD were no different from those in control patients (0.47 +/- 0.07 vs 0.45 +/- P =.08). Compared with control values, mitochondria from patients with PAD had a significantly lower acceptor control ratio (1.41 +/- 0.10 vs 2.90 +/- 0.20; P <.001).

CONCLUSION

Mitochondrial respiratory activity is abnormal in lower extremity skeletal muscle in patients with PAD. When considered in concert with the ultrastructural and enzymatic abnormalities previously documented in mitochondria of chronically ischemic muscle, these data support the concept of defective mitochondrial function as a pathophysiologic component of PAD.

摘要

目的

已证实外周动脉疾病（PAD）患者骨骼肌线粒体存在离散的形态、酶活性和功能变化。我们检测了9例晚期PAD患者（10条腿）和9例无PAD证据的对照患者（9条腿）腓肠肌中的线粒体呼吸情况。

方法

在含有皂素透皮肌束的氧电极池中，用Clark电极测定线粒体呼吸速率。肌肉样本取自腓肠肌的前内侧，位于胫骨结节远端10 cm处。线粒体呼吸速率以每毫克非胶原蛋白每分钟消耗的氧纳米原子数计算，在基线时（V(0)）、添加底物（苹果酸和谷氨酸；V(SUB)）后、添加二磷酸腺苷（ADP）后（V(ADP)）以及最后用苍术苷抑制腺嘌呤核苷酸转位酶后（V(AT)）进行测量。接受者控制率是线粒体整体功能的敏感指标，计算方法为添加ADP后的呼吸速率与用苍术苷抑制腺嘌呤核苷酸转位酶后的呼吸速率之比（V(ADP)/V(AT)）。

结果

PAD患者肌肉线粒体的呼吸速率在基线时与对照值无显著差异（0.31±0.06对0.55±0.12；P = 0.09），但PAD患者的V(sub)显著低于对照受试者（0.43±0.07对0.89±0.20；P < 0.05），V(ADP)也是如此（0.69±0.13对1.24±0.20；P < 0.05）。PAD患者经苍术苷抑制后的呼吸速率与对照患者无差异（0.47±0.07对0.45±；P = 0.08）。与对照值相比，PAD患者的线粒体接受者控制率显著降低（1.41±0.10对2.90±0.20；P < 0.001）。