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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Retrovirus-mediated transfer of an angiotensin type I receptor (AT1-R) antisense sequence decreases AT1-Rs and angiotensin II action in astroglial and neuronal cells in primary cultures from the brain.
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Retrovirus-mediated transfer of an angiotensin type I receptor (AT1-R) antisense sequence decreases AT1-Rs and angiotensin II action in astroglial and neuronal cells in primary cultures from the brain.

机译：逆转录病毒介导的血管紧张素I型受体（AT1-R）反义序列的转移降低了大脑原代培养的星形胶质细胞和神经元细胞中的AT1-Rs和血管紧张素II的作用。

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摘要

The AT1-R has been implicated in many cellular and physiological actions of angiotensin II (AII) in the brain. A retrovirus vector (LNSV) containing an AT1B-R antisense sequence (AT1B-AS) (termed LNSV-AT1B-AS) was constructed and used to determine the feasibility of using viral-mediated gene transfer to control AT1-Rs and AII actions in astroglial and neuronal cells in primary cultures from rat brain. Briefly, a 1.26-kb antisense sequence corresponding to nt -132 to +1128 of AT1-R cDNA was cloned into the LNSV vector, the vector was transfected into PA317 cells, and transfected cells were selected in G418. Incubation of brain cells with culture medium containing LNSV-AT1B-AS viral particles showed that AT1B-AS was integrated into the genome and transcribed in brain cells. This was associated with a significant decrease in AT1-Rs and in the AII-stimulated increase of c-fos mRNA, a measure of AT1-R function. These observations show that the AT1B-AS gene can be transferred into astroglial cells in cultureby LNSV and that such a transfer inhibits AT1-Rs and the AII stimulation of cellular activities. In addition, the usefulness of this approach to study AII-dependent pathophysiology in primary neuronal cultures from brain, in particular, is established.
机译：AT1-R与大脑中血管紧张素II（AII）的许多细胞和生理作用有关。构建了包含AT1B-R反义序列（AT1B-AS）（称为LNSV-AT1B-AS）的逆转录病毒载体（LNSV），并用于确定使用病毒介导的基因转移来控制AT1-Rs和AII作用的可行性大鼠脑原代培养物中的星形胶质细胞和神经元细胞。简而言之，将对应于AT1-R cDNA的nt -132至+1128的1.26kb反义序列克隆到LNSV载体中，将该载体转染到PA317细胞中，并在G418中选择转染的细胞。用含有LNSV-AT1B-AS病毒颗粒的培养基孵育脑细胞表明，AT1B-AS已整合到基因组中并在脑细胞中转录。这与AT1-Rs的显着减少和AII刺激的c-fos mRNA的增加有关，这是对AT1-R功能的一种衡量。这些观察结果表明，通过LNSV可以将AT1B-AS基因转移到培养的星形胶质细胞中，并且这种转移抑制了AT1-Rs和细胞活性的AII刺激。此外，建立了这种方法对研究特别是来自大脑的原始神经元培养物中依赖于AII的病理生理学的有用性。

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