miRNA was analyzed in an AD group (n = 23) and an age-matched control group (n = 23) using commercially available miRNA arrays from LC Sciences. Up-regulation of miRNA-146a coupled to down-regulation of CFH was observed in AD brain and in interleukin-1beta, Abeta42, and/or oxidatively stressed human neural (HN) cells in primary culture. Transfection of HN cells using an NF-kappaB-containing pre-miRNA-146a promoter-luciferase reporter construct in stressed HN cells showed significant up-regulation of luciferase activity that paralleled decreases in CFH gene expression. Treatment of stressed HN cells with the NF-kappaB inhibitor pyrollidine dithiocarbamate or the resveratrol analog CAY10512 abrogated this response. Incubation of an antisense oligonucleotide to miRNA-146a (anti-miRNA-146a; AM-146a) was found to restore CFH expression levels. These data indicate that NF-kappaB-sensitive miRNA-146a-mediated modulation of CFH gene expression may in part regulate an inflammatory response in AD brain and in stressed HN cell models of AD and illustrate the potential for anti-miRNAs as an effective therapeutic strategy against pathogenic inflammatory signaling.
miRNA-146a up-regulation in AD brain. A, merge of cy3/cy5 signals from a human micro-RNA panel (LC Sciences) indicates specific up-regulation of miRNA-146a (position A6; arrow). Levels of miRNA-9 (position A4), miRNA-132 (position A9), or miRNA-185 (position B6) showed no such changes (complete miRNA grid pattern available from LC Sciences). B, nylon membrane-bound DNA equivalents (10 μm) of three brain-enriched miRNAs and 5SRNA were probed with total 32P-radiolabeled miRNA fractions isolated from control or AD affected hippocampal CA1; representative Northern hybridization of select brain-enriched miRNAs reconfirmed up-regulation of miR-146a (arrows). Position A1, miRNA-9; A2, miRNA-132; B1, miRNA-146a; B2, 5SRNA. C, signals were quantified against internal 5SRNA levels in bar graph format. Data analysis of 23 control (CON) and 23 Alzheimer (ALZ) brains show a 3.3-fold increase of miRNA-146a expression in the neocortex (nctx) and a 2.3-fold increase in the hippocampus (hipp) over age-matched controls. *, significance over control p < 0.01 (ANOVA).
miRNA Microarray Service – LC Sciences provides a microRNA (miRNA) expression profiling service using microarrays based on our in-house developed µParaflo® technology platform. We have standard arrays for all mature miRNAs of all species available in the latest version of the miRBase database (Release 21, July 2014). Our service is comprehensive and includes sample labeling, array hybridization, image data processing and in-depth data analysis. Two-three weeks after receiving your total RNA samples, we’ll send you both the raw and fully analyzed data. [Learn more…]
Zhao Y, Lukiw WJ. (2008) An NF-kappaB-sensitive micro RNA-146a-mediated inflammatory circuit in Alzheimer disease and in stressed human brain cells. J Biol Chem 283(46), 31315-22. [article]