NGF mediates the neuroprotective effect of the β2-adrenoceptor agonist clenbuterol in vitro and in vivo:: evidence from an NGF-antisense study
NGF mediates the neuroprotective effect of the β2-adrenoceptor agonist clenbuterol in vitro and in vivo:: evidence from an NGF-antisense study
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DOI：
10.1016/s0197-0186(99)00032-7
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发表时间：
1999-07-01
影响因子：
4.2
通讯作者：
Krieglstein, J
中科院分区：
文献类型：
--
作者：
Culmsee, C;Semkova, I;Krieglstein, J
我们实验室以前的研究表明，β（2）-肾上腺素受体激动剂克仑特罗在体外和体内的神经保护作用是由于神经生长因子的合成增加而发生的。本研究的目的是评估磷酸化的神经生长因子寡脱氧核苷酸对克伦特罗在体外和体内的神经保护作用的影响。在原代培养的大鼠皮质星形胶质细胞中，观察到克伦特罗处理（1-100 μ M）后神经生长因子mRNA和蛋白水平的增加（对照的200-300%）。神经生长因子反义寡核苷酸（0.3-1 μ M，3天）使星形胶质细胞培养基中神经生长因子蛋白的含量浓度依赖性地降低至对照水平的20%。混合海马细胞培养基中的神经生长因子含量降低至接受载体或随机对照寡核苷酸的姐妹培养物的55%。在随机寡核苷酸（1 μ M，30小时）预处理的混合海马培养物中，克伦特罗（10 μ M）减少谷氨酸暴露（0.5 mM，1小时）后受损神经元的百分比为17%。谷氨酸孵育前用神经生长因子反义寡核苷酸（1 μ M）预处理30 h阻断了克伦特罗的保护作用。在体内，克伦特罗（0.01-0.1 mg/kg）剂量依赖性地减少大鼠永久性局灶性脑缺血模型的梗死体积。脑缺血前皮层注射神经生长因子反义寡核苷酸可阻断克伦特罗的脑保护作用，本研究结果表明，神经生长因子反义寡核苷酸是研究神经生长因子敲低效应的有效工具。通过使用神经生长因子反义寡核苷酸，我们可以证明，神经生长因子介导的β（2）-肾上腺素受体激动剂克伦特罗在体外和体内的神经保护作用。(C)1999 Elsevier Science Ltd.保留所有权利。
Previous studies in our laboratory suggested that neuroprotective effects of the beta(2)-adrenoceptor agonist clenbuterol in vitro and in vivo occurred due to enhanced synthesis of nerve growth factor. The aim of the present study was to evaluate the effects of a phosphothioated NGF oligodeoxynucleotide on neuroprotection by clenbuterol in vitro and in vivo.After clenbuterol treatment (1-100 mu M) an increase in nerve growth factor mRNA and protein levels (200-300% of control) was observed in primary cultures of rat cortical astrocytes. Nerve growth factor antisense oligonucleotide (0.3-1 mu M for 3 days) reduced the content of nerve growth factor protein in the medium of the astrocytes concentration-dependently to 20% of control level. Nerve growth factor content in the medium of mixed hippocampal cells was reduced to 55% of sister cultures receiving the vehicle or a random control oligonucleotide. In mixed hippocampal cultures pretreated with random oligonucleotide (1 mu M, 30 h), clenbuterol (10 mu M) reduced the percentage of damaged neurons after glutamate exposure (0.5 mM, 1 h) to 17%. Pretreatment with nerve growth factor antisense oligonucleotide (1 mu M) for 30 h before glutamate incubation blocked the protective effect of clenbuterol. In vivo, clenbuterol (0.01-0.1 mg/kg) reduced the infarct volume in a rat model of permanent focal cerebral ischemia dose-dependently. Nerve growth factor antisense oligonucleotides injected into the cortical tissue before ischemia abolished the cerebroprotective effect of clenbuterol.Our results indicate that the nerve growth factor antisense oligonucleotide presented in this study is a useful tool to investigate the effects of nerve growth factor knock down. By using the nerve growth factor antisense oligonucleotide we could demonstrate that nerve growth factor mediated the neuroprotective effects of the beta(2)-adrenoceptor agonist clenbuterol in vitro and in vivo. (C) 1999 Elsevier Science Ltd. All rights reserved.