Detection of tumor necrosis factor-α mRNA induction in ischemic brain tolerance by means of real-time polymerase chain reaction

被引:61
作者
Wang, XK [1 ]
Li, X [1 ]
Erhardt, JA [1 ]
Barone, FC [1 ]
Feuerstein, GZ [1 ]
机构
[1] SmithKline Beecham Pharmaceut, Dept Cardiovasc Pharmacol, King Of Prussia, PA 19406 USA
关键词
TNF alpha; ischemic tolerance; ischemic preconditioning; focal stroke; TaqMan; polymerase chain reaction;
D O I
10.1097/00004647-200001000-00004
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
A short duration of ischemia (i.e., ischemic pre conditioning) results in significant brain protection to subsequent severe ischemic insult. Because previous studies suggest that tumor necrosis factor-alpha (TNF-alpha) plays a role in both promoting ischemic damage and neuroprotection, the present work aimed to evaluate the expression of TNF-alpha mRNA in an established model of ischemic preconditioning using a transient 10-minute occlusion of the middle cerebral artery. Because the level of TNF-alpha mRNA expression in the brain was too low to be consistently detected by Northern technique, a real-time polymerase chain reaction method was applied to quantitate the absolute copy number of TNF-alpha transcript in rat brain after the preconditioning procedure. TNF-alpha mRNA was induced in the ipsilateral cortex as early as 1 hour (27 +/- 1 copies of mRNA per microgram of tissue compared to 11 +/- 3 copies in sham-operated samples) after preconditioning, reached a peak level at 6 hours (49 +/- 10 copies of transcript, n = 4, P < 0.01), and persisted up to 2 days. These data not only demonstrate the utility of real-time polymerase chain reaction for sensitive and accurate measurement of mRNA expression in normal and injured tissues but also suggest a potential role of TNF-alpha in the phenomenon of ischemic preconditioning.
引用
收藏
页码:15 / 20
页数:6
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