Translational Paradigms in Cerebrovascular Gene Transfer

Khurana, Vini G.; Meyer, Fredric B.

doi:10.1097/01.wcb.0000093324.07718.d4

Cited by 13 publications

(3 citation statements)

References 80 publications

(204 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…8,12 Caspase-3 activity was measured as described previously. 13 Measurements of Nitrate/Nitrite Levels and Nicotinamide-Adenine Dinucleotide Phosphate Oxidase Activity NO levels (in the form of nitrate/nitrite) in brain extracts were measured at day 3 after MCAO as described previously. 14 Reduced nicotinamideadenine dinucleotide phosphate (NADH) oxidase activity was measured by lucigenin-enhanced chemiluminescent detection of superoxide in a luminometer (TD-20/20, Turner Designs Inc).…”

Section: Western Blot Analyses Of Akt and Glycogen Synthetase Kinase-mentioning

confidence: 99%

Postischemic Brain Injury Is Exacerbated in Mice Lacking the Kinin B2 Receptor

et al. 2006

View full text Add to dashboard Cite

Abstract-Kallikrein cleaves low molecular weight kininogen to generate vasoactive kinins, which bind to the kinin B2 receptor, triggering a host of biological effects. Kallikrein gene delivery has been shown previously to reduce ischemia/reperfusioninduced cerebral infarction. In this study, we tested the hypothesis that the kinin B2 receptor plays a protective role in ischemic brain injury using kinin B2 receptor gene knockout (B2R-KO) mice subjected to middle cerebral artery occlusion (MCAO). The mortality rate and neurological deficit scores of B2R-KO mice (nϭ48) after MCAO were significantly increased compared with wild-type (WT) mice (nϭ40) when examined over a 14-day period. In addition, the infarct volume in B2R-KO mice was significantly larger than in WT mice at days 1 and 3 after MCAO. Similarly, apoptotic cells, detected by TUNEL labeling counterstained with propidium iodide, and caspase-3 activity in the ischemic brain increased significantly in B2R-KO mice at days 1 and 3 after MCAO. Furthermore, the accumulation of neutrophils in the ischemic brain of B2R-KO mice after MCAO increased when compared with WT mice and was associated with elevated tumor necrosis factor ␣ expression. These alterations in B2R-KO mice correlated with decreased NO levels, Akt, and glycogen synthase kinase-3␤ phosphorylation and increased nicotinamide-adenine dinucleotide oxidase activity. These results indicate that the kinin B2 receptor promotes survival and protects against brain injury by suppression of apoptosis and inflammation induced by ischemic stroke. The tissue kallikrein-kinin system has been shown previously to exert protective effects in heart after ischemic injury. 6,7 In addition, intracerebroventricular injection of the kallikrein gene has been shown to reduce cerebral infarct size and promote glial cell migration and survival after ischemic stroke. 8 The effects of kallikrein gene delivery were associated with enhanced NO formation, decreased oxidative stress, and activation of cell survival signaling pathways. Our previous studies have shown that the benefits of kallikrein gene delivery in the ischemic heart can be abolished by the administration of a kinin B2 receptor antagonist. 7 Therefore, the kinin B2 receptor may also play an important role in protecting the brain from ischemia/reperfusion (I/R) injury.To determine whether kallikrein/kinin provides neuroprotective effects against ischemic stroke via the kinin B2 receptor, we used wild-type (WT) and kinin B2 receptor knockout (B2R-KO) mice subjected to 90-minute occlusion of the middle cerebral artery followed by reperfusion. We then examined the potential effects of kinin B2 receptor on cell death and inflammatory cell accumulation in the ischemic brain. Our results showed that neurological deficit scores, infarct volume, and mortality rate were significantly elevated in B2R-KO mice in association with increased severity of apoptosis and inflammation in the ischemic brain. Methods Animals and TreatmentsMice (male and female, body weight 25 to 30 g), i...

show abstract

Section: Western Blot Analyses Of Akt and Glycogen Synthetase Kinase-mentioning

confidence: 99%

Postischemic Brain Injury Is Exacerbated in Mice Lacking the Kinin B2 Receptor

et al. 2006

View full text Add to dashboard Cite

show abstract

“…7,34,35 In the present study, we found that intracisternal protein transduction using an 11R-fusion protein selectively delivered this protein into cerebral vessels, and the delivered protein was especially transduced into the tunica media (smooth muscle layer) of the BA (Figure 3), even when it had been exposed to SAH (Figures 4, 5). This finding suggests that this protein transduction method may be a more effective therapeutic The high expression of 11R-EGFP was maintained when the BAs were kept on incubating with 11R-EGFP for 12 hours ex vivo (Figure 1).…”

Section: Discussionmentioning

confidence: 85%

Novel Protein Transduction Method by Using 11R

Ogawa

Ono

Ichikawa

et al. 2007

Stroke

View full text Add to dashboard Cite

Background and Purpose-A motif of 11 consecutive arginines (11R) is reported to be one of the most effective protein transduction domains for introducing proteins into the cell membrane. We therefore examined the transduction efficiency of 11R in cerebral arteries. Methods-Basilar arteries (BAs) obtained from rats were incubated with either 11R-enhanced green fluorescent protein (11R-EGFP) or EGFP without 11R. After incubation, expression of 11R-EGFP or EGFP in BA serial sections was observed by fluorescence microscope. In an additional in vivo experiment, 11R-EGFP or EGFP was injected into the cisterna magna with or without subarachnoid hemorrhage. The 11R-EGFP or EGFP was injected just after the autologous blood injection, and then the expression of 11R-EGFP or EGFP in BA sections was also observed by fluorescence microscope. Results-The 11R-EGFP signal was much stronger than that of EGFP in all layers of the rat BA, in both in vivo and ex vivo experiments. Moreover, the 11R-EGFP was transduced into the BA immediately (2 hours after the injection). Interestingly, 11R-fused fluorescent protein was transduced especially into the tunica media of the BA. Conclusions-The 11R-fused fluorescent protein effectively penetrates into all layers of the rat BA, especially into the tunica media. This is the first study to our knowledge to demonstrate the successful transduction of a protein transduction domain fused protein into the cerebral arteries.

show abstract

“…A literatura enfatiza que a patogênese do vasoespasmo cerebral tardio está relacionada com vários processos patológicos que incluem a destruição endotelial e A exposição da hemoglobina nas cisternas acaba sendo a mesma ligada hepaticamente e produzindo a haptoglobina (HP) que possui duas isoformas e está relativamente ligada ao processo da inflamação 186 .…”

Section: E2 Fisiopatologia Do Vasoespasmo Encefálicounclassified

Biomarcadores genéticos na hemorragia subaracnoidea aneurismática em pacientes da Amazônia

Paschoal¹

View full text Add to dashboard Cite

Translational Paradigms in Cerebrovascular Gene Transfer

Cited by 13 publications

References 80 publications

Postischemic Brain Injury Is Exacerbated in Mice Lacking the Kinin B2 Receptor

Postischemic Brain Injury Is Exacerbated in Mice Lacking the Kinin B2 Receptor

Novel Protein Transduction Method by Using 11R

Biomarcadores genéticos na hemorragia subaracnoidea aneurismática em pacientes da Amazônia

Contact Info

Product

Resources

About