改良St.Thomas灌注液预灌注对犬骨骼肌缺血再灌注损伤的保护  

Protective effect of pre-perfusion with improved St.Thomas solution on skeletal muscle ischemia-reperfusion injury in dogs

作　　者：赵磊 杨智伟 夏超 王国军 童梁成 杨俊生 汪剑龄 李颖 ZHAO Lei;YANG Zhiwei;XIA Chao;WANG Guojun;TONG Liangcheng;YANG Junsheng;WANG Jianling;LI Ying(Department of Orthopeadics,Air Force Hospital of Eastern Theater of PLA,Nanjing 210002,China)

机构地区：[1]中国人民解放军东部战区空军医院骨科,南京210002

出　　处：《中华显微外科杂志》2022年第6期656-663,共8页Chinese Journal of Microsurgery

基　　金：中国人民解放军后勤部卫生部科研基金(11MA059)。

摘　　要：目的探究改良St.Thomas灌注液预灌注对犬骨骼肌缺血再灌注损伤(IRI)的保护作用及机制。方法2021年3月-2021年9月,在解放军东部战区空军医院实验手术室,将16只比格犬通过随机数字表法分为对照组、IRI组、IRI加生理盐水(IRI+NS)组和改良St.Thomas组,每组4只,构建犬骨骼肌IRI模型,通过预灌注改良St.Thomas灌注液[氯化钾(KCl)、硫酸镁(MgSO4)以及NaHCO3(pH调整)],监测犬生命体征,通过苏木素-伊红(HE)染色、电镜检测和组织湿/干重比探究犬骨骼肌病理损伤,通过标记血管和低氧诱导因子-1α(HIF-1α)检测其血管密度和乏氧表现。构建L6鼠成肌细胞IRI模型,预孵育St.Thomas灌注液各组分,探究对细胞增殖抑制的影响,并通过检测还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)、F2异前列腺素(F2-isoprostane)、白介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、髓过氧化物酶(MPO)、谷胱甘肽过氧化物酶(GSH-Px)等探究其保护机制。采用统计软件SPSS 23.0进行统计学处理,P<0.05为差异有统计学意义。结果改良St.Thomas组犬生命体征较稳定,多巴胺维持量少,腓肠肌组织病理结构趋于完整,组织细胞及线粒体肿胀明显缓解,组织湿/干重比小于IRI组(P=0.046)。预孵育治疗剂量的MgSO4或NaHCO3,细胞增殖率大于IRI组(P<0.01、P=0.005),NADPH(P=0.004、P=0.01)、F2-isoprostane(P<0.001、P=0.01)、IL-1β(P=0.02、P=0.015)、TNF-α(P<0.01、P<0.01)、MPO(P<0.01、P<0.01)均较IRI组下降,GSH-Px较IRI组升高(P<0.01、P<0.001)。结论预灌注改良St.Thomas灌注液,可在短期内使犬生命体征趋于平稳,灌注液可能通过抗炎、抗氧化应激,改善骨骼肌细胞状态,改善组织缺氧,减轻骨骼肌组织细胞损伤。Objective To explore the protective effect and mechanism of improved St.Thomas solution on canine skeletal muscle ischemia-reperfusion injury(IRI).Methods Between March 2021 and September 2021,in the experimental operating room at the Air Force Hospital of the PLA Eastern Theater Command,16 Beagles were randomly divided into control group,IRI group,IRI+NS group,and improved St.Thomas group,4 in each group.The canine skeletal muscle IRI model was established,and the canine vital signs were monitored by pre-perfusion with improved St.Thomas perfusate[potassium chloride(KCl),magnesium sulfate(MgSO4),and NaHCO3(pH adjusted)].The pathological damage of canine skeletal muscle was explored by hematoxylin eosin(HE)staining,electron microscope detection and tissue wet/dry weight ratio,and blood vessel density.Hypoxia performances were detected by labeling blood vessels and hypoxia-inducible factor-1α(HIF-1α).The IRI model of L6 rat myoblasts was established,and the components of St.Thomas perfusion solution were pre incubated to explore the effect on the inhibition of cell proliferation.And by detecting reduced nicotinamide adenine dinucleotide phosphate(NADPH),F2 isoprostane(F2-isoprostane),interleukin 1β(IL-1β),tumour necrosis factor alpha(TNF-α),myeloperoxide enzyme(MPO),glutathione peroxidase(GSH-Px),etc.to explore its protective mechanism.Statistical software SPSS 23.0 was used for statistical analysis,A P<0.05 was set as statistically significant.Results In the improved St.Thomas group,the vital signs of the dogs were relatively stable,the amount of maintained dopamine was less,the histopathological structure of the gastrocnemius muscle tended to be intact,the swelling of tissue cells and mitochondria was significantly relieved,and the tissue wet/dry weight ratio was less than that in the IRI group(P=0.046).Pre-incubated with therapeutic doses of MgSO4 or NaHCO3,the proliferation rate of L6 cells was higher than that of IRI group(P<0.01,P=0.005),NADPH(P=0.004,P=0.001),F2-isoprostane(P<0.01,P=0.01),IL-1β(P=0.02

关 键 词：缺血再灌注损伤 骨骼肌 改良心脏停跳液 肢体损伤 犬 

分 类 号：R68[医药卫生—骨科学]