Evaluation of neuron damage H E staining Three days post TBI, each group of rats was sacrificed using an overdose of pentobarbital and then perfused transcardially with 0. 9% NaCl and 10% formalin. twice After perfusion, the rats were decapitated and their brains were removed and embedded in paraffin blocks. Coronal sections were stained with H E and subjected to microscopic examination. Evaluation of axonal demyelination Luxol fast blue staining Using a similar procedure to the one described above, the axonal damage was also analyzed. The embedded coronal Inhibitors,Modulators,Libraries sections were stained with luxol fast blue and cresyl echt violet for myelin detection and axonal loss assessment. Statistical analysis The obtained data are presented as the meansstandard error of the mean.
Kruskal Wallis analyses of variance were conducted, and if significant, were followed by Inhibitors,Modulators,Libraries the MannWhitney U test. P 0. 05 was considered statistically significant. Results Upregulation of Nogo A after TBI The first experiment conducted in the current study sought to examine alterations in the expression of Nogo A in the hippocampus after TBI. Compared with the sham group, the Nogo A mRNA expression level was found to rise slightly at four hours after TBI induction, but this difference was not significant. The upregulation of Nogo A expression reached a maximum at eight hours after trauma and lasted for three days. This stimulatory effect on Nogo A production was further con firmed by protein analysis. Western blot analysis revealed an increase in Nogo A protein in the hippocampus four hours post TBI.
However, a statistically significant elevation n the protein level Inhibitors,Modulators,Libraries began at eight hours after TBI and lasted for three days. Moreover, this TBI induced stimulation Inhibitors,Modulators,Libraries of Nogo A expression could be reversed by the administration of Nogo A antisense oligonucleotide immediately after TBI. As shown in the RT PCR analysis and western blot analysis, microinjection of Nogo A antisense oligonucleotide into the lateral ventricle drastically decreased the TBI induced Nogo A production by approximately 70%. However, the Nogo A irrelevant control oligonucleotide appeared to be ineffective in decreasing the TBI associated Nogo A production. Indomethacin attenuated expression of Nogo A Indomethacin, a potent non steroidal anti inflammatory drug, was used in this experiment to determine the relation ship between TBI associated inflammatory effects and Nogo A expression.
The level of Nogo A was again signifi cantly increased as a consequence Inhibitors,Modulators,Libraries of TBI, whereas in the TBI rats that were given indomethacin, Nogo A expres sion at both the mRNA and protein levels returned to those observed selleck screening library in sham animals. Unlike the direct effect conferred by Nogo A antisense oligonucleotide, indomethacin may conceivably have triggered a novel pathway that resulted in the sup pression of Nogo A expression.
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