目的 研究安菲博肽(ANF)对大鼠心肌缺血再灌注损伤的保护作用及其作用机制。
方法 取SD大鼠96只,随机分为正常组,模型组,地尔硫
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组,安菲博肽高、中、低(4、2、1 μg·kg
-1)组,每组16只。腹腔注射给药连续3 d后,结扎大鼠左冠状动脉前降支30 min再灌注120 min 建立心肌缺血再灌注损伤模型。氯化三苯基四氮唑 (TTC)染色测定心肌梗死面积;HE染色观察心肌组织病理改变;生化试剂盒测定血清中肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)活性,ELISA法测定血清中心肌肌钙蛋白I(cTnI)、白介素1β(IL-1β)、白介素6(IL-6)、肿瘤坏死因子α(TNF-α)含量;Western-blot法测定心肌组织中Toll样受体4(TLR4)、髓样分化因子(MyD88)、核因子κB(NF-κB p65/p-NF-κB p65)蛋白表达。
结果 与模型组比较,ANF高、中剂量组可明显减少心肌梗死面积,改善心肌组织病理改变;高、中、低剂量组可明显降低血清中CK-MB、LDH活性及cTnI、IL-6含量;高、中剂量组可明显降低血清中IL-1β、TNF-α含量以及心肌组织中TLR4、MyD88、p-NF-κB蛋白表达水平。
结论 ANF对大鼠心肌缺血再灌注损伤具有明显的保护作用,其机制可能与抑制TLR4/NF-κB信号通路激活从而减轻炎症反应有关。
Abstract
OBJECTIVE To study the effect and mechanism of anfibatide(ANF) on myocardial ischemia reperfusion injury in rats. METHODS Ninety-six SD rats were randomly divided into six groups: sham, model, positive (Dil 10 mg·kg-1) and ANF 4, 2,1 μg·kg-1 group. After 3 d of intraperitoneal injection, myocardial ischemia reperfusion injury model was established by ligating the left anterior descending cornary artery for 30 min and reperfuion for 120 min. Using TTC and HE staining to determine the myocardial infarction area and the tissue pathological change. The activities of creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH) in serum were measured by biochemical kit. Serum levels of cardiac troponin I (cTnI), interleukin 1β(IL-1β), interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA). Protein expressions of toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and nuclear factor κB (NF-κB p65/p-NF-κB p65) were determined by Western-blot assay. RESULTS Compared with model group, ANF could significantly reduce cardiac I/R-induced the infarct size and histological damage in rat myocardium, markedly inhibite the increase of LDH and CK-MB activities and cTnI, IL-1β, IL-6 and TNF-α levels in rats subjected to myocardial I/R injury. In addition, ANF could obviously prevent myocardial I/R-increased TLR4, MyD88 and p-NF-κB protein expressions. CONCLUSION ANF has protective effect on myocardial I/R injury in rats, which maybe related to inhibiting the activation of TLR4/NF-κB signaling pathway and thereby reducing the inflammatory response.
关键词
安菲博肽 /
心肌缺血再灌注 /
炎症反应 /
TLR4-NF-κB信号通路
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Key words
anfibatide /
myocardial ischemia reperfusion /
inflammatory response /
TLR4/NF-κB signaling pathway
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中图分类号：
R965
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脚注
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基金
安徽省自然科学基金资助(1608085MH161)
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