右美托咪定减轻氧化应激改善H9C2细胞缺氧/复氧损伤(1)
摘 要：目的 探讨右美托咪定减轻氧化应激改善H9C2细胞缺氧/复氧损伤的作用。方法 H9C2心肌细胞随机分为三组，正常对照组(Control)、缺氧/复氧组(H/R)、Dex(5 μmol/L)干预H/R组。Dex干预H/R组先用Dex预处理6 h后，再经缺氧/复氧处理。检测各组细胞培养基中LDH的含量，CCK-8法检测各组H9C2心肌细胞的存活率，试劑盒检测caspase-3活性，试剂盒检测MDA的含量和SOD活性。结果 与Control组相比，H/R组细胞活力降低，培养基中LDH含量增加，caspase-3活性增加，MDA含量升高而SOD活性降低，统计学意义显著(P<0.01)；Dex预处理提高H/R处理后的细胞活力，减少LDH含量和caspase-3活性，降低MDA含量，增加SOD活性，统计学意义显著(P<0.01)。结论 Dex可通过减轻氧化应激改善H9C2心肌细胞缺氧/复氧损伤。
关键词：右美托咪定；氧化应激；缺氧/复氧；H9C2
中图分类号：R614 文献标识码：A DOI：10.3969/j.issn.1006-1959.2018.14.027
, http://www.100md.com
文章编号：1006-1959(2018)14-0095-03
Abstract：Objective To investigate the effect of dexmedetomidine on oxidative stress and improvement of hypoxia/reoxygenation injury in H9C2 cells.Methods H9C2 cardiomyocytes were randomly divided into three groups.The normal control group(Control)， hypoxia/reoxygenation group(H/R)，and Dex(5μmol/L)were used to intervene in the H/R group.Dex intervention in the H/R group was pretreated with Dex for 6 h and then treated with hypoxia/reoxygenation.The LDH content in the cell culture medium was detected. The survival rate of H9C2 cardiomyocytes in each group was detected by CCK-8 method.The caspase-3 activity was detected in the kit， and the MDA content and SOD activity were detected by the kit.Results Compared with the Control group，the cell viability in the H/R group decreased，the LDH content in the medium increased，the caspase-3 activity increased，the MDA content increased and the SOD activity decreased，statistically significant(P<0.01)；Dex pretreatment improved cell viability after H/R treatment，decreased LDH content and caspase-3 activity，decreased MDA content，and increased SOD activity，with statistical significance(P<0.01).Conclusion Dex can alleviate hypoxia/reoxygenation injury of H9C2 cardiomyocytes by reducing oxidative stress.
, 百拇医药
Key words：Dexmedetomidine；Oxidative stress；Hypoxia/Reoxygenation；H9C2
缺血性心肌病(Ischemic cardiomyopathy，ICM)是世界范围内具有高致残率和致死率的疾病，是目前威胁人类生命健康的一类主要疾病[1]。快速有效的实现缺血心肌的血液复流即再灌注，是临床上首选的治疗方法，然而再灌注本身会加重心肌损伤，导致能量代谢障碍，形成心肌缺血再灌注损伤(myocardial ischemia reperfusion injury， MIRI)[2-4]。关于MIRI机制的研究集中在钙超载，氧化应激，能量代谢障碍，炎症反应和凋亡、坏死及自噬等方面[5-7]。右美托咪定(dexmedetomidine， Dex)是一种新型的高选择性α2受体激动剂，具有剂量依赖性的镇静、镇痛、抗焦虑和抑制交感神经兴奋等作用，且对呼吸、循环的抑制作用轻微[8]。Dex对心肌的保护作用受到越来越多的关注，普遍认为Dex是通过抑制交感中枢活动，降低心率，降低心肌氧耗，改善心肌氧供需平衡，但具体机制仍未阐明。本研究采用H9C2细胞缺氧/复氧模型，观察Dex预处理减轻氧化应激改善H9C2细胞缺氧/复氧损伤的作用。
1材料与方法
1.1试剂与仪器 H9C2心肌细胞系购买自深圳百恩维公司；盐酸右美托咪定购自江苏恒瑞医药有限公司(200 μg/2 ml)；DMEM培养液和胎牛血清购自美国Gibco公司；0.25 %胰蛋白酶购自Sigma公司；CCK-8细胞毒性检测试剂盒购自七海生物公司；LDH检测试剂盒，caspase-3活性检测试剂盒，MDA检测试剂盒和SOD检测试剂盒均购自南京建成生物工程研究所。超净工作台购自苏州净化设备仪器厂；CO2细胞培养箱购自Thermo公司；低速离心机购自赛特湘仪公司；酶标仪购自SpectraMax公司。, http://www.100md.com(李璟)
关键词：右美托咪定；氧化应激；缺氧/复氧；H9C2
中图分类号：R614 文献标识码：A DOI：10.3969/j.issn.1006-1959.2018.14.027
, http://www.100md.com
文章编号：1006-1959(2018)14-0095-03
Abstract：Objective To investigate the effect of dexmedetomidine on oxidative stress and improvement of hypoxia/reoxygenation injury in H9C2 cells.Methods H9C2 cardiomyocytes were randomly divided into three groups.The normal control group(Control)， hypoxia/reoxygenation group(H/R)，and Dex(5μmol/L)were used to intervene in the H/R group.Dex intervention in the H/R group was pretreated with Dex for 6 h and then treated with hypoxia/reoxygenation.The LDH content in the cell culture medium was detected. The survival rate of H9C2 cardiomyocytes in each group was detected by CCK-8 method.The caspase-3 activity was detected in the kit， and the MDA content and SOD activity were detected by the kit.Results Compared with the Control group，the cell viability in the H/R group decreased，the LDH content in the medium increased，the caspase-3 activity increased，the MDA content increased and the SOD activity decreased，statistically significant(P<0.01)；Dex pretreatment improved cell viability after H/R treatment，decreased LDH content and caspase-3 activity，decreased MDA content，and increased SOD activity，with statistical significance(P<0.01).Conclusion Dex can alleviate hypoxia/reoxygenation injury of H9C2 cardiomyocytes by reducing oxidative stress.
, 百拇医药
Key words：Dexmedetomidine；Oxidative stress；Hypoxia/Reoxygenation；H9C2
缺血性心肌病(Ischemic cardiomyopathy，ICM)是世界范围内具有高致残率和致死率的疾病，是目前威胁人类生命健康的一类主要疾病[1]。快速有效的实现缺血心肌的血液复流即再灌注，是临床上首选的治疗方法，然而再灌注本身会加重心肌损伤，导致能量代谢障碍，形成心肌缺血再灌注损伤(myocardial ischemia reperfusion injury， MIRI)[2-4]。关于MIRI机制的研究集中在钙超载，氧化应激，能量代谢障碍，炎症反应和凋亡、坏死及自噬等方面[5-7]。右美托咪定(dexmedetomidine， Dex)是一种新型的高选择性α2受体激动剂，具有剂量依赖性的镇静、镇痛、抗焦虑和抑制交感神经兴奋等作用，且对呼吸、循环的抑制作用轻微[8]。Dex对心肌的保护作用受到越来越多的关注，普遍认为Dex是通过抑制交感中枢活动，降低心率，降低心肌氧耗，改善心肌氧供需平衡，但具体机制仍未阐明。本研究采用H9C2细胞缺氧/复氧模型，观察Dex预处理减轻氧化应激改善H9C2细胞缺氧/复氧损伤的作用。
1材料与方法
1.1试剂与仪器 H9C2心肌细胞系购买自深圳百恩维公司；盐酸右美托咪定购自江苏恒瑞医药有限公司(200 μg/2 ml)；DMEM培养液和胎牛血清购自美国Gibco公司；0.25 %胰蛋白酶购自Sigma公司；CCK-8细胞毒性检测试剂盒购自七海生物公司；LDH检测试剂盒，caspase-3活性检测试剂盒，MDA检测试剂盒和SOD检测试剂盒均购自南京建成生物工程研究所。超净工作台购自苏州净化设备仪器厂；CO2细胞培养箱购自Thermo公司；低速离心机购自赛特湘仪公司；酶标仪购自SpectraMax公司。, http://www.100md.com(李璟)