Koca Kenan, Yurttaş Yüksel, Yıldız Cemil, Caycı Tuncer, Uysal Bülent, Korkmaz Ahmet
GATA, Department of Orthopedics, Ankara, Turkey.
Acta Orthop Traumatol Turc. 2010;44(6):476-83. doi: 10.3944/AOTT.2010.2327.
The aim of the study was to investigate the effect of hyperbaric oxygen-preconditioning (HBO-PC) and ozone-preconditioning (O3-PC) on oxidative/nitrosative stress induced by tourniquet ischemia/reperfusion (I/R) in rat skeletal muscle.
Thirty-two Wistar-Albino-type male rats included in the study were divided into four groups of equal number: 1) sham operation, 2) I/R, 3) I/R+HBO-PC, or 4) I/R+O3-PC. One session of 3-4 L/min 100% oxygenation for 60 min at 3 absolute atmosphere (ATA) was defined as one dose of HBO; in total, 7 doses of HBO-PC were administered before ischemia. One dose of O3 comprised 0.7 mg/kg ozone/oxygen mixture, administered intraperitoneally; a total of 4 doses of O3-PC were administered before ischemia. The I/R model was performed in anesthetized rats by clipping right femoral artery to induce 2 h ischemia followed by 22 h of reperfusion. The right gastrocnemius muscle and venous blood samples were harvested. Tissue was assayed for levels of malondialdehyde (MDA), inducible nitric oxide synthase (iNOS), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). Serum was assayed to measure the level of nitrite-nitrate (NOx).
Tissue MDA level, SOD activity, and serum NOx level were increased, whereas GSHPx was decreased in the I/R group. MDA and NOx levels were decreased, whereas GSH-Px activity was increased in both the I/R+HBO-PC and I/R+O3-PC groups. SOD activity was increased in the I/R+O3-PC group, but did not change significantly in the I/R+HBO-PC group. iNOS staining score and intensity were lower in the I/R+HBO-PC and I/R+O3-PC groups than I/R group.
Both O3-PC and HBO-PC reduced tissue lipid peroxidation, NOx levels, and iNOS staining scores in the experimental I/R model. Our data suggest that HBO-PC and O3-PC protect against oxidative/nitrosative stress induced by I/R in rat skeletal muscle.
本研究旨在探讨高压氧预处理（HBO-PC）和臭氧预处理（O3-PC）对大鼠骨骼肌止血带缺血/再灌注（I/R）诱导的氧化/亚硝化应激的影响。
纳入研究的32只Wistar白化型雄性大鼠被分为数量相等的四组：1）假手术组，2）I/R组，3）I/R+HBO-PC组，或4）I/R+O3-PC组。在3个绝对大气压（ATA）下以3-4 L/min的速度进行60分钟的100%氧合作为1剂HBO；在缺血前共给予7剂HBO-PC。1剂O3包括0.7 mg/kg臭氧/氧气混合物，经腹腔注射；在缺血前共给予4剂O3-PC。通过夹闭右股动脉在麻醉大鼠中建立I/R模型，诱导2小时缺血，随后再灌注22小时。采集右腓肠肌和静脉血样本。检测组织中丙二醛（MDA）、诱导型一氧化氮合酶（iNOS）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）的水平。检测血清中亚硝酸盐-硝酸盐（NOx）水平。
I/R组组织MDA水平、SOD活性和血清NOx水平升高，而GSHPx降低。I/R+HBO-PC组和I/R+O3-PC组的MDA和NOx水平降低，而GSH-Px活性升高。I/R+O3-PC组SOD活性升高，但I/R+HBO-PC组无显著变化。I/R+HBO-PC组和I/R+O3-PC组的iNOS染色评分和强度低于I/R组。
在实验性I/R模型中，O3-PC和HBO-PC均降低了组织脂质过氧化、NOx水平和iNOS染色评分。我们的数据表明，HBO-PC和O3-PC可保护大鼠骨骼肌免受I/R诱导的氧化/亚硝化应激。
