Al-Jasmi Fatma, Pramathan Thachillath, Swid Adnan, Sahari Bahjat, Penefsky Harvey S, Souid Abdul-Kader
Department of Paediatrics, College of Medicine & Health Sciences, United Arab Emirates University, Al Ain, United Arab Emirates;
Sultan Qaboos Univ Med J. 2013 Aug;13(3):411-6. doi: 10.12816/0003264. Epub 2013 Jun 25.
This study investigated the feasibility of using a phosphorescence oxygen analyser to measure cellular respiration (mitochondrial O2 consumption) in foreskin samples and their fibroblast-rich cultures.
Foreskin specimens from normal infants were collected immediately after circumcision and processed for measuring cellular respiration and for culture. Cellular mitochondrial O2 consumption was determined as a function of time from the phosphorescence decay of the Pd (II) meso-tetra-(4-sulfonatophenyl)-tetrabenzoporphyrin.
In sealed vials containing a foreskin specimen and glucose, O2 concentration decreased linearly with time, confirming the zero-order kinetics of O2 consumption by cytochrome oxidase. Cyanide inhibited O2 consumption, confirming that the oxidation occurred mainly in the mitochondrial respiratory chain. The rate of foreskin respiration (mean ± SD) was 0.074 ± 0.02 μM O2 min(-1) mg(-1) (n = 23). The corresponding rate for fibroblast-rich cultures was 9.84 ± 2.43 μM O2 min(-1) per 10(7) cells (n = 15). Fibroblast respiration was significantly lower in a male infant with dihydrolipoamide dehydrogenase gene mutations, but normalised with the addition of thiamine or carnitine.
The foreskin and its fibroblast-rich culture are suitable for assessment of cellular respiration. However, the clinical utility of foreskin specimens to detect disorders of impaired cellular bioenergetics requires further investigation.
本研究调查了使用磷光氧分析仪测量包皮样本及其富含成纤维细胞的培养物中细胞呼吸(线粒体氧气消耗)的可行性。
正常婴儿的包皮标本在包皮环切术后立即收集,并进行细胞呼吸测量和培养处理。通过钯(II)中-四-(4-磺基苯基)-四苯并卟啉的磷光衰减来确定细胞线粒体氧气消耗随时间的变化。
在含有包皮标本和葡萄糖的密封小瓶中,氧气浓度随时间呈线性下降,证实了细胞色素氧化酶消耗氧气的零级动力学。氰化物抑制氧气消耗,证实氧化主要发生在线粒体呼吸链中。包皮呼吸速率(平均值±标准差)为0.074±0.02μM O₂ min⁻¹ mg⁻¹(n = 23)。富含成纤维细胞的培养物的相应速率为每10⁷个细胞9.84±2.43μM O₂ min⁻¹(n = 15)。一名患有二氢硫辛酰胺脱氢酶基因突变的男婴的成纤维细胞呼吸明显较低,但添加硫胺素或肉碱后恢复正常。
包皮及其富含成纤维细胞的培养物适用于评估细胞呼吸。然而,包皮标本在检测细胞生物能量受损疾病方面的临床实用性需要进一步研究。