Role for Metallothioneins-I/II in Isoflurane Preconditioning of Primary Murine Neuronal Cultures
Role for Metallothioneins-I/II in Isoflurane Preconditioning of Primary Murine Neuronal Cultures
复制标题
DOI：
10.1097/aln.0b013e3181974bba
复制
发表时间：
2009-03-01
期刊：
影响因子：
8.8
通讯作者：
Hall, Adam C.
中科院分区：
文献类型：
--
作者：
Edmands, Scott D.;Hall, Adam C.
背景：用吸入麻醉剂（包括异氟烷）进行预处理，可以在包括脑组织在内的多个组织中诱导持久的细胞保护，抵抗缺血引起的毒性。金属调节基因金属硫蛋白-I/II (MT-I/II) 已被证明可以保护多种组织免受氧化损伤。此外，还发现 MT 对异氟烷和缺血预处理的反应存在差异性调节。在本研究中，我们评估了 MT-I/II 在介导原代神经元胶质细胞培养物中异氟烷预处理中的作用。方法：在 3 小时氧-葡萄糖剥夺（OGD，缺血模型）前 24-96 小时用异氟烷（3 小时，1.5%）对原代小鼠神经元胶质细胞进行预处理。 OGD 后，通过乳酸脱氢酶释放评估异氟烷保护以及 MT-I/II 敲低和敲除培养物对预处理的反应性。微管相关蛋白 2 和胶质纤维酸性蛋白的免疫测定确定了神经元-胶质细胞对预处理的敏感性。通过定量逆转录酶-聚合酶链反应评估 MT-I/II 信使 RNA。分析用外源 MT-I/II 转染的培养物针对 OGD 毒性的保护作用。结果：异氟醚预处理在 24 小时时将 OGD 介导的毒性降低了 1-1.6 +/- 7.9%，预处理后保护作用在 72 It 时增加到 37.5 +/- 2.5%。免疫标记显示，与神经胶质细胞相比，神经元对 OGD 更敏感，对异氟烷预处理更敏感。定量逆转录酶-聚合酶链式反应显示，异氟烷处理后 MT-I/II 信使 RNA 上调（约 2.5 倍）。 MT-I/II 蛋白转染也显着降低了 OGD 介导的毒性。最后，MT-I/II 的敲除和敲除分别减弱和消除了异氟烷介导的保护。结论：MT-I/II 在异氟烷介导的体外神经元和神经胶质细胞 OGD 毒性的延迟预处理中发挥着重要作用。
Background: Pretreatment with inhaled anesthetics, including isoflurane, can induce long-lasting cellular protection against ischemia-derived toxicity in multiple tissues, including brain tissue. Metal-regulatory genes, metallothioneins-I/II (MT-I/II), have been shown to protect against oxidative damage in multiple tissues. Furthermore, MT have been found to be differentially regulated in response to isoflurane and ischemic preconditioning. In this study, we assess the role of MT-I/II in mediating isoflurane preconditioning in primary neuronal-glial cultures.Methods: Primary mouse neuronal-glial cultures were preconditioned with isoflurane (3 h, 1.5%) 24-96 h before 3-h oxygen-glucose deprivation (OGD, ischemic model). After OGD, isoflurane protection and responsiveness of MT-I/II knockdown and knockout cultures to preconditioning were assessed by lactate dehydrogenase release. Immunoassays for microtubule associated protein 2 and glial fibrillary acidic protein determined neuronal-glial sensitivity to preconditioning. MT-I/II messenger RNA was assessed by quantitative reverse transcriptase-polymerase chain reaction. Cultures transfected with exogenous MT-I/II were analyzed for protection against OGD toxicity.Results: Isoflurane preconditioning reduced OGD-mediated toxicity by 1-1.6 +/- 7.9% at 24 h, with protection increasing to 37.5 +/- 2.5% at 72 It after preconditioning. Immunolabeling showed that neurons were more sensitive to OGD and more responsive to isoflurane preconditioning compared to glia. Quantitative reverse transcriptase-polymerase chain reaction showed MT-I/II messenger RNA were upregulated (approximately 2.5-fold) by isoflurane treatments. Also MT-I/II protein transfection significantly decreased OGD-mediated toxicity. Finally, knockdown and knockout of MT-I/II diminished and abolished isoflurane-mediated protection, respectively.Conclusions: MT-I/II play all important role in isoflurane-mediated delayed preconditioning against OGD toxicity of neuronal and glial cells in vitro.