血红素加氧酶-1在七氟醚预处理减轻乳鼠心肌细胞缺氧复氧损伤中的作用  

Role of heme oxygeuase-1 in process of sevoflurane preconditioning attenuating hypoxia-reoxygeuation injury in cultured neonatal rat cardiomyocytes

作　　者：孙志鹏[1] 罗婷[1] 黄德樱[1] 乐江[2] 

机构地区：[1]武汉市妇女儿童医疗保健中心麻醉科,430016 [2]武汉大学医学院药理学教研室

出　　处：《中华麻醉学杂志》2011年第8期1001-1004,共4页Chinese Journal of Anesthesiology

摘　　要：目的探讨血红素加氧酶-1（HO-1）在七氟醚预处理减轻乳鼠心肌细胞缺氧复氧损伤中的作用。方法新生健康清洁级SD大鼠15只，日龄1—3d，处死后取心室肌组织，原代培养心肌细胞，以1×10^6个／ml接种于6孔培养板或以2×10^5个／ml接种于24孔培养板，采用随机数字表法，将其随机分为4组（n=25）：对照组（C组）常规培养；缺氧复氧组（H／R组）采用缺氧2h，复氧1h的方法制备心肌细胞缺氧复氧损伤模型；七氟醚预处理组（S＋H／R组）细胞经2．5％七氟醚预处理20min后行药物洗脱10min，再行缺氧复氧处理；锌原卟啉＋七氟醚预处理组（ZnPP＋S＋H／R组）细胞经HO-1抑制剂锌原卟啉3／Lmol／L孵育1h后，行七氟醚预处理及缺氧复氧处理。于复氧结束后测定心肌细胞HO-1表达、细胞凋亡率、细胞内游离Ca^2＋浓度（[Ca^2＋]i）、线粒体膜通透性转运孔（m）开放程度、细胞色素C（Cyto C）表达及培养液LDH和CK活性。结果与C组比较，H／R组心肌细胞HO．1和胞浆CytoC表达上调，线粒体CytoC表达下调，培养液LDH、CK活性、细胞凋亡率、[Ca^2＋]i和PTP开放度升高（P〈O．01）。与H／R组比较，S＋H／R组心肌细胞HO．1和线粒体CytoC表达上调，胞浆CytoC表达下调，培养液LDH、CK活性、细胞凋亡率、[Ca^2＋]i和PTP开放度降低（P〈0．01）。与S＋H／R组比较，ZnPP＋s＋H／R组心肌细胞HO一1和线粒体CytoC表达下调，胞浆CytoC表达上调，培养液LDH、CK活性、细胞凋亡率、[Ca^2＋]i和肿开放度升高（P〈0．01）。结论HO-1表达上调参与了七氟醚预处理减轻乳鼠心肌细胞缺氧复氧损伤。Objective To investigate the role of heme oxygenase-1 （HO-1） in process of sevoflurane pre- conditioning attenuating hypoxia-reoxygenation （H/R） injury in cultured neonatal rat cardiomyocytes. Methods Primary cultured neonatal rat cardiomyocytes were randomly divided into four groups （ n = 25 each） ： control group （group C）, group H/R, sevoflurane preconditioning group（group S ＋ H/R）, and HO-1 inhibitor zinc protoporphyria （ZnPP） and sevoflurane preconditioning group （group ZnPP ＋ S ＋ H/R）. In group H/R, the cardiomyocytes were exposed to 2 hours of hypoxia, followed by 1 hour of reoxygenation. Group S ＋ H/R received 2.5% sevoflurane preconditioning for 20 minutes followed by 10 minutes of wash-out before H/R. ZnPP was added to the culture medium with final concentrations of 3 μmol/L 1 hour before sevoflurane preconditioning and H/R in group ZnPP ＋ S＋ H/R. HO-1 expression, apoptosis rate, concentration of free calcium（ [Ca^2＋] i）, mitochondrial membrane permeability transition pore （ PTP）, cytochrome C （ Cyto C） expression and activities of lactate dehydrogenase （LDH） and creatine kinase （CK） in culture supernatant were detected after reoxygenation. Results Compared with group C, the expression of HO-1 and cytoplasmic Cyto C of cardiomyocytes were up-regulated, mitochondrial Cyto C was down-regulated, while the [ Ca^2＋]i, opening degree of PTP, apoptosis rate and activities of LDH and CK in culture supernatant were increased in group H/R. Compared with group H/R, the expression of HO-1 and mitochondrial Cyto C of cardiomyocytes were upzregulated, cytoplasmic Cyto C was down-regulated, while the [Ca^2＋]i, opening degree of PTP, apoptosis rate and activities of LDH and CK in culture supernatant were decreased in group S ＋ H/R. Compared with group S ＋ H/R, the expression of HO-1 and mitochondrial Cyto C of cardiomyocytes were down-regulated, cytoplasmic Cyto C was up-regulated, while the [ Ca^2＋ ] i, opening degree of PTP, apoptosis rate and

关 键 词：血红素加氧酶.1 细胞低氧 氧 肌细胞 心脏 七氟醚 

分 类 号：R965[医药卫生—药理学]