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. 2012 May 3;23(5):1218–1229. doi: 10.1093/cercor/bhs115

Figure 3.

Figure 3.

CPAI prevents LPS/HI-induced chemokine production and monocyte recruitment. (A) Representative results of a cytokine array using brain extracts at 24 h following indicated conditions (n = 2 for unchallenged and LPS-treated animals; n = 4 for LPS/HI-PBS or LPS/HI-CPAI treatments). The duplicated spots of MCP-1 and the quantification results were indicated. LPS/HI increased the brain MCP-1 level to 9.75 ± 0.54 (mean ± SD) fold of the baseline level, while CPAI treatment attenuated this induction. (B and C) ELISA analysis of the MCP-1 levels from brains (B) or the plasma (C) at 24 h after the indicated conditions (n = 4 for each). The MCP-1 level in the unchallenged brain was 4.4 ± 0.85 pg/mL extracts (mean ± SD), which was increased by LPS/HI insults to 50.3 ± 4.4 pg/mL (asterisk, P < 0.01) and significantly attenuated by the CPAI treatment (P < 0.05 by unpaired t-test). In contrast, the basal plasma MCP-1 level was already high in unchallenged animals (116.7 ± 3.3 pg/mL), and upregulated by various stimuli to a similar degree. (D–I) Immunofluoresence detection of Iba1 (D, E, and F) and OX42/CD11b (G, H, and I) in coronal sections at the fornix-decussation level of brains collected at 16 h after pure-HI, LPS/HI-PBS, or LPS/HI-CPAI treatments (n> 5). White lines mark the boundaries of corpus callosum (CC). LV, lateral ventricles. Scale bar: 50 μm. Note that the CC in LPS/HI-PBS-treated rats was 2 times wider than that of LPS/HI-CPAI-treated rats and filled with numerous large, round Iba1/OX42 double-positive cells (248 ± 24 per visual field). In contrast, LPS/HI-CPAI-treated animals contained fewer Iba1+ cells within the corpus callosum (50 ± 11 per visual field) that exhibited ramified cytoplasmic processes and did not express OX42. Also note that the swelling of CC was milder in pure-HI-injured animal brains and there was no obvious OX42-immunoreactivity. (J) The tensor trace map from rat brain at 24 h after LPS/HI insult. Note the expansion/swelling of CC/external capsule on the HI-injured hemisphere (indicated by arrows).