Apolipoprotein E-/- mice have delayed skeletal muscle healing after hind limb ischemia-reperfusion
Apolipoprotein E-/- mice have delayed skeletal muscle healing after hind limb ischemia-reperfusion
复制标题
DOI：
10.1016/j.jvs.2008.04.006
复制
发表时间：
2008-09-01
影响因子：
4.3
通讯作者：
Watkins, Michael T.
中科院分区：
文献类型：
--
作者：
Kang, Jeanwan;Albadawi, Hassan;Watkins, Michael T.
引言：肢体缺血再灌注损伤的经典研究已使用年轻健康小鼠进行。然而，外周血管疾病患者年龄较大，经常表现出代谢紊乱，可能会延迟血运重建后的愈合。载脂蛋白E（ApoE（-/-））基因缺失的小鼠已被用作各种高胆固醇血症实验方案中的模型。这些实验评估了炎症反应和骨骼肌形态学的变化，在急性和慢性阶段的肢体缺血再灌注损伤ill aged ApoE（-/-）juice.Methods：ApoE（-/-）和野生型（Wt）小鼠进行了1.5小时的单侧后肢缺血，随后1，7，或14天的再灌注（DR）。骨骼肌纤维损伤的组织学分析在DR时进行评估。肌纤维成熟的形态学证据在14 DR时进行评估。在7和14 DR时，使用蛋白质印迹法评估骨骼肌分化标志物MyoD和肌细胞生成素的水平。炎症标志物，包括髓过氧化物酶，巨噬细胞炎性蛋白-2（MIP-2），单核细胞趋化蛋白-1（MCP-1）和骨桥蛋白，采用酶联免疫吸附试验和趋化因子（C-C基序）受体2（CCR-2）在1，7和14 DR采用蛋白质印迹法进行测定。在1DR后，测量组织腺苷5 '-三磷酸（ATP）水平以评估代谢活性。结果：1DR后骨骼肌组织学检查显示，Wt和ApoE（-/-）小鼠骨骼肌损伤程度无显著性差异。然而，在14 DR时，ApoE（-/-）小鼠具有比Wt小鼠更高的未成熟肌纤维百分比。在7 DR时，ApoE（-/-）小鼠中的肌细胞生成素水平较低。ApoE-/-小鼠的损伤骨骼肌在7 DR时的髓过氧化物酶水平低于Wt小鼠，在14 DR时的MCP-1水平高于Wt小鼠。组织中ATP、MIP-2、骨桥蛋白或CCK 2的水平在所有实验intervention.Conclusion：虽然Wt和ApoE（-/-）小鼠在再灌注急性期的损伤肌肉之间没有差异，但ApoE-/-小鼠在再灌注慢性期的骨骼肌愈合延迟。在ApoE（-/-）小鼠中，肌肉再生的这种滞后与7 DR时肌细胞生成素水平较低和14 DR时MCP-1水平升高有关。ApoE（-/-）小鼠骨骼肌愈合延迟可能对具有血管风险因素（如高胆固醇血症）的老年患者的组织愈合不良和功能恢复具有更广泛的意义。
Introduction: Classic studies of limb ischemia-reperfusion injury have been performed using young healthy mice. However, patients with peripheral vascular disease are older and often exhibit metabolic derangements that may delay healing after revascularization. Mice with genetic deletion of apolipoprotein E (ApoE(-/-)) have been used as a model ill various experimental scenarios of hypercholesterolemia. These experiments evaluated the inflammatory response and changes in skeletal muscle morphology during the acute and chronic phases of limb ischemia-reperfusion injury ill aged ApoE(-/-) juice.Methods: Age-matched ApoE(-/-) and wild-type (Wt) mice underwent 1.5 hours of unilateral hind limb ischemia, followed by 1, 7, or 14 days of reperfusion (DR). Histologic analysis of skeletal muscle fiber injury was assessed at I DR. Morphologic evidence of muscular fiber maturation was assessed at 14DR. Levels of MyoD and myogenin, markers of skeletal muscle differentiation, were assessed at 7 and 14DR using Western blots. Markers of inflammation, including myeloperoxidase, macrophage inflammatory protein-2 (MIP-2), monocyte chemotactic protein-1 (MCP-1), and osteopontin, were assayed using enzyme-linked immunosorbent assay and chemokine (C-C motif) receptor 2 (CCR-2) using Western blots at 1, 7, and 14DR. After 1DR, tissue adenosine 5'-triphosphate (ATP) levels were measured to assess metabolic activity. Unpaired t test and Mann-Whitney test were used for comparisons.Results: Histologic evaluation of skeletal muscle after 1DR showed no difference in the degree of injury between Wt and ApoE(-/-) mice. However, at 14DR, ApoE(-/-) mice had higher percentage of immature muscle fibers than Wt mice. Myogenin level was lower in the ApoE(-/-) mice at 7DR. Injured skeletal muscle of ApoE-/- mice had lower levels of myeloperoxidase than Wt mice at 7 DR and higher levels of MCP-1 at 14DR. There was no difference in the levels of tissue ATP, MIP-2, osteopontin, or CCK2 at all experimental intervals.Conclusion: Although there was no difference between the injured muscle of Wt and ApoE(-/-) mice during the acute phase of reperfusion, ApoE-/- mice showed delay in skeletal muscle healing during the chronic phase of reperfusion. This lag in muscle regeneration was associated with lower levels of myogenin at 7DR and an increased level of MCP-1 at 14DR in the ApoE(-/-) mice. The delay in skeletal muscle healing in the ApoE(-/-) mice may have broader implications for poor tissue healing and functional recovery in elderly patients who have vascular risk factors such as hypercholesterolemia.