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. 2013 Jan 9;33(3):396–406. doi: 10.1038/jcbfm.2012.179

Figure 4.

Figure 4

Knockdown of SirT1 by short interfering RNA (siRNA) reverses neuroprotection of hyperbaric oxygen (HBO) preconditioning in rat brain. (A, B) Representative western blot bands and quantitative evaluation of SirT1 protein expression (n=5/group). Rats were treated with a single intracerebroventricular (i.c.v.) injection of 5 μg siRNA-SirT1 or siRNA-C. The expression of SirT1 was detected by immunoblots from day 1 to day 7 after siRNA injection (A). At 24 hours before HBO preconditioning, 5 μg siRNA-SirT1 or siRNA-C was delivered by a single i.c.v. injection. The expression of SirT1 was detected by immunoblots 24 hours after reperfusion (B). The expression of SirT1 was normalized to the expression of β-actin. Data are presented as mean±s.d., one-way analysis of variance (ANOVA) followed by the least significant difference (LSD) test. (C) Neurobehavioral score 24 hours after reperfusion (n=8/group). Data are presented as the median (range), a nonparametric method (Kruskal–Wallis test) followed by the Mann–Whitney U-test by Bonferroni correction. (D) Representative 2,3,5-triphenyltetrazolium chloride-stained thick brain sections and quantitative evaluation of the infarct volume 24 hours after reperfusion (n=8/group). Data are presented as mean±s.d., one-way ANOVA followed by (LSD test. *P<0.01 versus middle cerebral artery occlusion (MCAO); #P<0.01 versus HBO preconditioning. siRNA-SirT1, SirT1 short interfering RNA; siRNA-C, control short interfering RNA. MCAO: rats subjected to occlusion of middle cerebral artery for 120 minutes; HBO+MCAO: rats subjected to MCAO 24 hours after the end of HBO preconditioning.