P-Glycoprotein Exacerbates Brain Injury Following Experimental Cerebral Ischemia by Promoting Proinflammatory Microglia Activation

Figure 6

P-glycoprotein silence or overexpress in endothelial cells regulates CCL2 expression and microglia phenotype following oxygen glucose deprivation/reoxygenation. Endothelial cells (bEnd.3) were transfected with P-glycoprotein (P-gp) or negative control (NC) siRNA, P-gp or NC pcDNA3.1 plasmid, or untransfected, and then subjected to either oxygen glucose deprivation/reoxygenation (OGD/R) treatment or normal culture conditions. Following 24 hr coculture with microglia (BV2), cells were harvested for RT-PCR assay and medium was collected for ELISA assay. (a, c) mRNA expression levels of CD16, iNOS, CD206, Arg-1 (in BV2 cells), CCL2, and CCR2 (in bEnd.3 cells) measured via RT-PCR assay as fold changes relative to control treatment (n = 4). (b, d) Contents of CD16, iNOS, CD206, Arg-1, and CCL2 in Transwell systems determined by ELISA assay (n = 3). Mann–Whitney test for (a) and (c). One-way ANOVA followed by the post hoc least significant difference test or Games Howell test for (b) and (d). All data are mean ± SD; , between two groups.

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