Ekinci Şafak, Kaldırım Ümit, Akyıldız Faruk, Bilgiç Serkan, Koca Kenan, Poyrazoğlu Yavuz, Uysal Ozgür Selim, Turğut Hasan, Türkkan Selim, Erşen Ömer, Topal Turgut, Ozkan Huseyin
Department of Orthopaedics and Traumatology, Ağrı Military Hospital, Ağrı, Turkey.
Department of Emergency Medicine, Gulhane Military Medical Academy, Ankara, Turkey.
Open Med (Wars). 2015 Feb 27;10(1):194-200. doi: 10.1515/med-2015-0031. eCollection 2015.
The aim of this study was to investigate the effect of hypothermia (H) on skeletal ischemia-reperfusion (IR) injury in rats by measuring malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), nitric oxide (NO), and interleukin-1 beta (IL-1β) in muscle, and measureing immunohistochemical-inducible nitric oxide synthase (iNOS) staining of skeletal muscle.
Eighteen Wistar Albino rats were divided randomly into three groups (sham, IR, hypothermia) (n=6). The sham group had all procedures without the IR period. The lower right extremity of rats in the IR and hypothermia groups was subjected to 2 hours of ischemia and 22 hours of reperfusion by applying a clamp on the common iliac artery and a rubber-band at the level of the lesser trochanter under general anesthesia. Rats in the hypothermia group underwent 4 hours of hypothermia during the first four hours of reperfusion in addition to a 2-hour ischemia and 22-hour reperfusion period. All rats were sacrificed at end of the IR period using a high dose of anesthesia. The tibialis anterior muscles were preserved. Immunohistochemical iNOS staining was performed, and MDA, SOD, GSH-Px, NO, and IL-1β were measured in the muscle.
The level of MDA, NO, and IL-1β in muscle was increased in the IR group compared with that in the sham group, but these parameters were decreased in the hypothermia group compared with the IR group. The activities of SOD and GSH-Px in muscle were decreased in the IR group; however, these parameters were increased in the hypothermia group. The score and intensity of iNOS staining of skeletal muscle was dens in IR group, mild in hypothermia group, and weak in sham group.
The present study has shown that hypothermia reduced IR injury in the skeletal muscle by decreasing the levels of MDA, NO, and IL-1β, and increasing the activities of SOD and GSH-Px. In addition, hypothermia attenuated the score and intensity of iNOS staining.
本研究旨在通过测量肌肉中的丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、一氧化氮（NO）和白细胞介素-1β（IL-1β），并检测骨骼肌免疫组化诱导型一氧化氮合酶（iNOS）染色，来研究低温（H）对大鼠骨骼肌缺血再灌注（IR）损伤的影响。
18只Wistar白化大鼠随机分为三组（假手术组、IR组、低温组）（n = 6）。假手术组进行所有操作，但无IR期。IR组和低温组大鼠在全身麻醉下通过在髂总动脉上夹闭和在小转子水平扎橡皮筋，使右下肢缺血2小时，再灌注22小时。低温组大鼠除了2小时缺血和22小时再灌注期外，在再灌注的前4小时进行4小时低温处理。在IR期结束时，使用高剂量麻醉处死所有大鼠。保留胫前肌。进行免疫组化iNOS染色，并测量肌肉中的MDA、SOD、GSH-Px、NO和IL-1β。
与假手术组相比，IR组肌肉中MDA、NO和IL-1β水平升高，但与IR组相比，低温组这些参数降低。IR组肌肉中SOD和GSH-Px活性降低；然而，低温组这些参数升高。骨骼肌iNOS染色的评分和强度在IR组中密集，在低温组中轻度，在假手术组中较弱。结论：本研究表明，低温通过降低MDA、NO和IL-1β水平，增加SOD和GSH-Px活性，减轻了骨骼肌的IR损伤。此外，低温减弱了iNOS染色的评分和强度。
