腺病毒介导的一氧化氮合酶基因转移可增加大鼠脑血流量。

Adenoviral gene transfer of nitric oxide synthase increases cerebral blood flow in rats.

作者信息

Lüders J C, Weihl C C, Lin G, Ghadge G, Stoodley M, Roos R P, Macdonald R L

机构信息

Section of Neurosurgery, Pritzker School of Medicine, University of Chicago Medical Center, Illinois 60637, USA.

出版信息

Neurosurgery. 2000 Nov;47(5):1206-14; discussion 1214-5. doi: 10.1097/00006123-200011000-00039.

DOI:10.1097/00006123-200011000-00039

PMID:11063115

Abstract

OBJECTIVE

Depletion of nitric oxide may play a role in the development of vasospasm after aneurysmal subarachnoid hemorrhage. Replenishment of nitric oxide might be a useful treatment for vasospasm. Using rats, we performed intracisternal injections of replication-defective adenovirus containing the endothelial nitric oxide synthase (eNOS) gene and determined the localization of and effect on cerebral blood flow of transgene expression.

METHODS

Rats underwent baseline measurement of cortical cerebral blood flow using laser Doppler flowmetry. Replication-defective adenovirus containing the Escherichia coli LacZ gene (Ad327beta-Gal, n = 2/time point) or the bovine eNOS gene (AdCD8-NOS, n = 4/time point) or physiological saline solution was injected into the cisterna magna. Cerebral blood flow was measured 1, 2, 4, 7, or 14 days later, and the animals were killed. Expression of beta-galactosidase activity from the LacZ gene was examined by histochemical staining and that of eNOS was examined by polymerase chain reaction assays of messenger ribonucleic acid. Brains were histopathologically examined for inflammation.

RESULTS

Beta-galactosidase activity was observed throughout the leptomeninges and in some cells in the adventitia of small subarachnoid blood vessels in the Ad327beta-Gal-injected rats. Messenger ribonucleic acid for eNOS was detected in the leptomeninges and brainstem 1 and 2 days after injection of AdCD8-NOS. Rats injected with Ad327beta-Gal or physiological saline solution exhibited decreased cerebral blood flow beginning 2 days after virus injection and lasting up to 14 days after injection. Rats injected with AdCD8-NOS developed significant transient increases in cerebral blood flow 2 days after virus injection, followed by slight decreases in blood flow. There was inflammation in the subarachnoid space of all animals; the inflammation was qualitatively worse in animals injected with Ad327beta-Gal, compared with rats injected with AdCD8-NOS or saline solution.

CONCLUSION

Intracisternal injection of replication-defective adenovirus containing the eNOS gene can transiently increase cerebral blood flow.

摘要

目的

一氧化氮耗竭可能在动脉瘤性蛛网膜下腔出血后血管痉挛的发生中起作用。补充一氧化氮可能是治疗血管痉挛的一种有效方法。我们利用大鼠进行脑池内注射含内皮型一氧化氮合酶（eNOS）基因的复制缺陷型腺病毒，并确定转基因表达的定位及其对脑血流的影响。

方法

使用激光多普勒血流仪对大鼠进行皮质脑血流的基线测量。将含大肠杆菌LacZ基因的复制缺陷型腺病毒（Ad327β - Gal，每个时间点n = 2）、含牛eNOS基因的复制缺陷型腺病毒（AdCD8 - NOS，每个时间点n = 4）或生理盐水注入大鼠脑池。在注射后1、2、4、7或14天测量脑血流，然后处死动物。通过组织化学染色检测LacZ基因的β - 半乳糖苷酶活性表达，通过信使核糖核酸的聚合酶链反应分析检测eNOS的表达。对大脑进行组织病理学检查以评估炎症情况。

结果

在注射Ad327β - Gal的大鼠中，在整个软脑膜以及蛛网膜下腔小血管外膜的一些细胞中观察到β - 半乳糖苷酶活性。注射AdCD8 - NOS后1天和2天，在软脑膜和脑干中检测到eNOS的信使核糖核酸。注射Ad327β - Gal或生理盐水的大鼠在病毒注射后2天开始脑血流降低，并持续至注射后14天。注射AdCD8 - NOS的大鼠在病毒注射后2天脑血流出现显著短暂增加，随后血流略有下降。所有动物的蛛网膜下腔均有炎症；与注射AdCD8 - NOS或生理盐水的大鼠相比，注射Ad327β - Gal的动物炎症在性质上更严重。