Tsui J C S, Baker D M, Shaw S G, Shi-Wen X, Dashwood M R
Department of Surgery, Royal Free and University College Medical School, London, UK.
Br J Surg. 2008 Jan;95(1):72-9. doi: 10.1002/bjs.5910.
Dysfunction of the nitric oxide pathway is implicated in peripheral arterial disease. Nitric oxide synthase (NOS) isoforms and NOS activity were studied in muscle from patients with critical leg ischaemia (CLI). Alterations in NOS during revascularization surgery were also assessed.
Muscle biopsies were taken from patients with CLI undergoing amputation and also from patients undergoing femorodistal bypass at the start of surgery, after arterial clamping and following reperfusion. The presence of NOS within muscle sections was confirmed using reduced nicotinamide adenine dinucleotide phosphate diaphorase histochemistry. NOS isoform distribution was studied by immunohistochemistry. NOS mRNA and protein levels were measured using real-time reverse transcriptase-polymerase chain reaction and western blotting. NOS activity was assessed with the citrulline assay.
All three NOS isoforms were found in muscle, associated with muscle fibres and microvessels. NOS I and III protein expression was increased in CLI (P = 0.041). During revascularization, further ischaemia and reperfusion led to a rise in NOS III protein levels (P = 0.008). NOS activity was unchanged.
Alterations in NOS I and III occurred in muscle from patients with CLI and further changes occurred during bypass surgery.
一氧化氮途径功能障碍与外周动脉疾病有关。对严重下肢缺血（CLI）患者的肌肉中一氧化氮合酶（NOS）亚型和NOS活性进行了研究。还评估了血管重建手术过程中NOS的变化。
在截肢的CLI患者以及在手术开始时、动脉夹闭后和再灌注后的股腘动脉搭桥患者中采集肌肉活检样本。使用还原型烟酰胺腺嘌呤二核苷酸磷酸黄递酶组织化学法确认肌肉切片中NOS的存在。通过免疫组织化学研究NOS亚型分布。使用实时逆转录聚合酶链反应和蛋白质印迹法测量NOS mRNA和蛋白质水平。用瓜氨酸测定法评估NOS活性。
在肌肉中发现了所有三种NOS亚型，它们与肌纤维和微血管相关。CLI患者中NOS I和III蛋白表达增加（P = 0.041）。在血管重建过程中，进一步的缺血和再灌注导致NOS III蛋白水平升高（P = 0.008）。NOS活性未发生变化。
CLI患者的肌肉中NOS I和III发生了改变，并且在搭桥手术过程中进一步发生了变化。
