骨骼肌缺血后调适动态pH测量及模拟酸灌注减轻缺血再灌注损伤

发布时间：2018-04-28 17:00

本文选题：再灌注损伤 + 肌　；参考：《中国组织工程研究》2017年16期

【摘要】：背景:研究报道,心肌缺血后调适的保护是通过心肌再灌注初期维持3 min的酸中毒起到作用,而在骨骼肌方面是否有此作用尚不清楚。目的:对大鼠骨骼肌缺血后调适过程进行动态pH测量,并模拟缺血后调适pH变化配置酸性灌注液输注,检测其对缺血再灌注损伤的影响。方法:基于前期大鼠骨骼肌缺血再灌注损伤建模方法和缺血后调适方案,运用光纤pH仪于主缺血期、4个循环30 s再灌/30 s缺血后调适操作期及其后再灌注期对大鼠骨骼肌进行pH连续测量,并用乳酸及生理盐水配置与缺血后调适期等pH灌注液备用。将25只健康成年雄性SD大鼠随机分为假手术组、缺血再灌注组、缺血后调适组、乳酸酸灌注组、生理盐水组5组,每组5只。各组按其实验方案进行相应实验并抽血检测乳酸脱氢酶,取样腓肠肌测算湿/干质量比值、髓过氧化物酶及TTC染色测算梗死面积,取样右侧胫前肌行Western Blot检测MAPK通路关键蛋白Erk1/2的表达。结果与结论:(1)缺血后调适于再灌注初期出现一个延长酸性平台,pH为6.81±0.133,时长为2 min 40 s。(2)乳酸脱氢酶、髓过氧化物酶、湿/干质量比值检测结果显示:缺血后调适组、乳酸酸灌注组明显低于缺血再灌注组(P0.05);(3)Western Blot分析显示:p-Erk的表达缺血后调适组、乳酸酸灌注组及生理盐水组均显著高于缺血再灌注组(P0.05);(4)TTC染色检测显示,缺血后调适组及乳酸酸灌注组梗死面积较缺血再灌注组明显减少(P0.05);(5)结论:缺血后调适可维持再灌注初期短暂酸性状态;酸性灌注液可以有效模拟缺血后调适作用减少大鼠骨骼肌缺血再灌注损伤。其机制可能通过Erk1/2磷酸化激活RISK信号通路而起到保护作用。
[Abstract]:Background: it is reported that the protective effects of myocardial ischemia and adaptation are due to the maintenance of acidosis for 3 min at the beginning of myocardial reperfusion, but it is not clear whether this effect can be achieved in skeletal muscle. Aim: to measure the dynamic pH of skeletal muscle after ischemia in rats, and to investigate the effect of acid infusion on ischemia reperfusion injury by simulating the change of pH after ischemia. Methods: based on the model of ischemic reperfusion injury of skeletal muscle in rats and the adjustment scheme after ischemia. The pH of skeletal muscle of rats was continuously measured by optical fiber pH meter during the main ischemic period, 4 circulatory 30 s reperfusion operation period and 30 s post-ischemia operation period and then reperfusion period. The pH was prepared with lactic acid and physiological saline configuration and after ischemia adjustment period. Twenty-five adult male SD rats were randomly divided into sham-operation group, ischemia-reperfusion group, post-ischemic conditioning group, lactic acid perfusion group, physiological saline group, 5 rats in each group. Lactate dehydrogenase was detected by blood sampling, wet / dry weight ratio was measured by sampling gastrocnemius muscle, infarct area was measured by myeloperoxidase and TTC staining. The expression of Erk1/2, the key protein of MAPK pathway, was detected by Western Blot in the right tibial anterior muscle. Results and conclusion: a prolonged acidic plateau appeared at the early stage of reperfusion. The pH was 6.81 卤0.133, and the duration was 2 min 40 s. 2) lactate dehydrogenase, myeloperoxidase and wet / dry weight ratio were detected. Western Blot analysis showed that the expression of p-Erk in lactic acid group was significantly lower than that in ischemia reperfusion group. The expression of p-Erk in lactic acid perfusion group and normal saline group was significantly higher than that in ischemia reperfusion group. The infarct size of the ischemic conditioning group and lactic acid perfusion group was significantly less than that of the ischemia reperfusion group. Conclusion: the adjustment after ischemia can maintain the transient acidic state in the early stage of reperfusion. Acid perfusion solution can effectively simulate the ischemic conditioning effect to reduce skeletal muscle ischemia reperfusion injury in rats. The mechanism may play a protective role by activating the RISK signaling pathway by Erk1/2 phosphorylation.
【作者单位】：广西医科大学第一附属医院创伤骨科手外科;
【基金】：国家自然科学基金资助项目(81260276)~~
【分类号】：R68

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