目的 研究4-羟基-2,2,6,6-四甲基哌啶基-N-氧化物(tempol)对高原缺氧小鼠心肌组织的保护作用及机制。方法 将110只小鼠随机分为正常对照组、缺氧模型组、乙酰唑胺组和tempol组,单次腹腔注射给药30 min后,在模拟海拔8 000 m环境停留12 h,眼眶取血后,测定血清中乳酸脱氢酶(LDH)和肌酸激酶(CK)活性,然后处死小鼠,检测心肌组织中过氧化氢(H2O2)和丙二醛(MDA)含量以及ATP酶和抗氧化酶的活性,蛋白印迹法检测缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)、核因子E2相关因子2(Nrf2)和血红素氧合酶-1(HO-1)蛋白的表达水平。结果 与正常对照组相比,缺氧模型组血清中CK和LDH的活性显著增加,心肌组织中H2O2和MDA含量显著升高,ATP酶和抗氧化酶的活性显著降低,HIF-1α、VEGF、Nrf2和HO-1蛋白表达增强。经tempol预处理后能够显著降低高原缺氧小鼠血清中CK和LDH的活性,减少心肌组织中H2O2和MDA含量,提高ATP酶和抗氧化酶的活性,降低HIF-1α、VEGF蛋白表达,显著提高Nrf2和HO-1蛋白表达。结论 Tempol对高原缺氧诱导的心肌组织损伤具有保护作用,该作用与改善能量代谢,清除自由基,激活Nrf2/HO-1信号途径,提高抗氧化酶活性,降低机体氧化应激有关。
Abstract
OBJECTIVE To study the effect mechanism of tempol against hypobaric hypoxia-induced heart damage in mice. METHODS One hundred and ten BALB/c mice were randomly divided into normal control group, hypoxia model group, acetazolamide group and tempol group. After single intraperitoneal injection for 30 min, the mice were exposed to a simulated high altitude of 8 000 m for 12 h. After hypoxic exposure, blood was collected from the eye sockets and separated into serum to measure the activities of lactic dehydrogenase (LDH)and creatine kinase (CK). Then the mice were sacrificed and the content of H2O2 and malondialdehyde (MDA) as well as ATPase and antioxidant enzyme activity in heart were determined. HIF-1, VEGF, Nrf2, and HO-1 were detected by immunohistochemistry. RESULTS Compared with normal control group, the activities of plasma CK and LDH in hypoxia model group significantly increased. In addition, the content of H2O2 and MDA in hypoxia model group significantly increased while ATPase and antioxidant enzymes activity markedly decreased compared with the normal control group. Moreover, the expression of HIF-1α, VEGF, Nrf2 and HO-1 increased. Prior administration of tempol effectively decreased the activities of plasma CK and LDH as well as the content of H2O2 and MDA in heart tissue. Tempol could increase ATPase and antioxidant enzyme activities and decreased the expression of HIF-1α and VEGF compared with hypoxia model, while it could further increase the expression of Nrf2 and HO-1. CONCLUSION Tempol has protective effect on heart injury induced by hypobaric hypoxia in mice. Its mechanism may be attributed to the amelioration of energy metabolism, scavenging free radical, improvement of antioxidant enzyme activity the activation of the Nrf2/HO-1 pathway as well as alleviation of oxidative stress.
关键词
4-羟基-2 /
2 /
6 /
6-四甲基哌啶基-N-氧化物 /
低压低氧 /
心肌损伤 /
抗氧化酶 /
Nrf2/HO-1途径
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Key words
tempol /
hypobaric hypoxia /
heart injury /
antioxidant enzyme /
Nrf2/HO-1 pathway
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中图分类号：
R965.1
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参考文献
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脚注
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基金
国家自然科学基金资助项目(81202458);全军医药科研“十二五”面上项目资助(CLZ12JA04);甘肃省自然科学基金资助项目(1308RJYA06);中国博士后科学基金资助项目(2012M521926)
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