Figure 2
CBD promotes a pattern of anti-inflammatory and anti-oxidative stress-related gene expression. Brain regions from both vocal-motor (HVC and RA) and -learning circuits (Area X) were micropunch dissected, total RNA extracted, cDNA synthesized, and PCR amplified. Gene expression of IL-1B, IL-6, IL-10, TNFα, and SOD2 were normalized to the endogenous control (GAPDH) and fold change from the unlesioned hemisphere was expressed as 2−∆∆CT. To quantify, cDNA from n = 5 per group was amplified in triplicate and the average cycle threshold (Ct) value was calculated. The mean Ct value was then used for further analysis. (A) CBD significantly decreased mean fold expression of pro-inflammatory IL-6 in HVC, RA, and Area X. (B) CBD decreased mean fold expression of IL-1Β in HVC and RA but not Area X. (C) TNFα was decreased in HVC and RA but not Area X. (D) mean fold expression of the anti-inflammatory mediator IL-10 was increased in HVC and RA but not significantly in Area X. (E) Expression of the marker of oxidative stress, superoxide dismutase 2 (SOD2) was decreased within HVC and RA. Note that tissue was obtained 24 h post-lesion and thus represents a “snap-shot” of inflammatory cytokine expression that is known to vary with time post-injury24. Group differences were assessed by mixed-models ANOVA with Sidak’s multiple comparison correction.
