乌司他丁对大鼠骨骼肌缺血-再灌注损伤的保护作用

Protective effects of ulinastalin on ischemla- reperfusion injury of skeletal muscle in rats

目的探讨乌司他丁对大鼠骨骼肌缺血-再灌注损伤的保护作用。方法24只健康成年雄性SD大鼠随机分为3组（每组8只）：对照组（C组）、缺血-再灌注损伤组（I／R组）、乌司他丁处理组（U组）。C组仅麻醉肢体没有缺血操作；I／R组于再灌注前颈外静脉注入生理盐水0．5ml；U组于再灌注前同法给乌司他丁0．5ml（5×10^4／kg）。以橡皮带环绕结扎大鼠左后肢根部至趾掌无血流信号达4h后放开橡皮带，再灌注4h建立大鼠骨骼肌缺血-再灌注损伤模型。再灌注4h各组处死动物采集标本，采用逆转录-聚合酶链反应（RT-PCR）法测定骨骼肌组织TNF-α mRNA的表达；酶联免疫吸附试验法（ELISA）测定血浆TNF-α水平；比色法检测血浆乳酸脱氢酶（LDH）、肌酸激酶（CK）、丙二醛（MDA）和组织过氧化物酶（咖的）含量，测定湿质量／干质量均（W／D）及光镜、电镜观察骨骼肌组织结构的变化。应用SPSS10．0统计软件，用单因素方差分析。结果骨骼肌组织TNF-α mRNA的表达在I／R组明显高于C组（P〈0．05），而U组则低于I／R组（P〈0．01）；血浆俐Ra浓度I／R组明显高于C组（P〈0．01），而U组则低于I／R组（P〈0．05）；血浆LDH、MDA、CK和组织MDA、MPO水平以及W／D比值I／R组高于C组（P〈0．05），U组则低于I／R组（P〈0．05）；骨骼肌组织形态学和超微结构的损伤U组也较I／R组减轻。结论乌司他丁通过抑制细胞因子的生成、抑制氧自由基代谢产物MDA的产生和减少MPO的活性而对骨骼肌缺血，再灌注起保护作用。

Objective To investigate the protective effects of ulinastatin （UTI） on ischemia-repeffusion （I/ R） injury （SMIRI） of skeletal muscle in rots. Method Twenty - four male SD rats randomly were divided into three groups in equal number： control group （Group C） rats underwent anesthetization without ischemia; the ischemia-repeffusion injury group （Group I/R） rats underwent ischemia and reperfusion, 0.5 ml normal saline （N. S） was infused upon extermal jugular vein prior to reperfusion; ulinstatin group （Group U） underwent ischemia and reperfusion, and 0.5 ml UT/ （5 × 10^4 U/kg） was infused at the same time. The skeletal muscle injury model was induced by a rubber band tourniquet applied to the left root of the hind limb for 4 hours and reperfusion for 4 hours. At the end of study, the expression of TNF-α mRNA of skeletal muscle was determined by reverse transcription-polymerase chain reaction （RT-PCR） ; enzyme-linked immtm-osorbent assay （ELISA） were performed for plasma level of TNF-α; the plasma concentrations of lactate dehydrogenase （LDH）, creatine kinase （CK）, malondialdehyde （MDA）, myeloperoxidase （MT） activity and MDA in skeletal muscle were measured by colorimetry respectively. The oedema degree was quantified by calculating the wet/dry weight ratio of skeletal muscle. Structural changes of skeletal muscle were also observed histologically and ultmstructurally. The statistical difference was analyzed with One-way ANOVA by SPSS version 10.0 Software for windows. Results The level of plasma TNF-α, TNF-α mRNA expression in skeletal muscle in group I/R were significantly higher than those in Group C （P 〈 0.01 ）, while those in Group U were significantly lower than those in Group I/R （P 〈 0.01 ）. The plasma concentrations of LDH, CK, MDA and the MPO activity, W/D of skeletal muscle varied in those groups were likewise in comparinson between groups （ P 〈 0.05）. The histologic changes of skeletal muscle tissue under light and electronic microscopy were slingter in Group U than in group I/R. Conclusions UTI can inhibit the production of inflammatory factors and MDA, and suppress the MPO activity, showing protective effects against ischemia-reperfusion injury of skeletal muscle of rats.