Despite the surge of interest in miRNA identification and quantification during the epileptogenic Acetylcorynoline process. Ideally, reference genes should present high expression stability levels in different experimental conditions. The evaluation of a panel of five candidate reference genes to Dimesna determine the most reliable one for accurate normalization of gene expression in the systemic PILO-model indicated twoas the most stable in the hippocampus of rats from all experimental and control groups. Depending on the software used, the rank of these genes on a stability scale was slightly different, probably because of the different mathematical algorithm employed. We also considered whether selecting multiple reference genes in combination is better than selecting a single reference gene alone. The optimal number of reference genes which should be used for accurate normalization was determined by calculating the normalization factor. The use of more than the two most stable reference genes identifiedis not required as suggested by the V-value. In order to evaluate the functional significance of the results obtained for reference genes, we conducted a relative expression analysis of the miR-146a gene, whose pattern is already described for the PILO-injected model. miR-146a can be induced by different pro-inflammatory stimuli and has been shown to be upregulated in experimental models of epilepsy, as well as in human TLE. Indeed, in PILO and electrical stimulation TLE models, prominent upregulation of miRNA-146a was evident at 13 week after SE and persisted in the chronic phase. In human TLE with hippocampal sclerosis, increased astroglial expression of miR-146a was observed mainly in regions where neuronal cell loss and reactive gliosis occurred. Similarly, Iyer et al.showed an overexpression of mir-146a in epilepsyassociated glioneuronal lesions. It remains unclear how the induction of mir146 expression may contribute to the etiopathogenesis of TLE. Thus, Iyer et al.observed that seizures alone may not account for changes in miR-146a expression. Moreover, emerging data suggest that miR-146a is induced as a negativefeedback regulator of the glial-mediated inflammatory response associated to the epileptogenic process. This is in line with other studies supporting an immunomodulatory role ascribed to miR-146a in several human neurodegenerative diseases associated with a strong inflammatory component. In fact, upregulation of miR146a has been detected in active multiple sclerosis lesions, in human Alzheimer diseasebrainand in prion disease, indicating an underlying common inflammatory response to different neurological insults. Accordingly, when normalized using the best combination of two or three reference genes, we observed that miR-146a transcript levels were significantly increased in the chronic stage. Interestingly, under our experimental conditions, the use of a reference gene individually for normalization leads to the relative transcript levels of the mir-146a gene to be different from those obtained with the best combinations of genes, and hence probably less accurate. This suggests, therefore, that an appropriate normalization strategy for miRNA expression during epileptogenesis requires the use of two or more reference genes. Curiously, in previous studies, under the same experimental conditions, in order to normalize protein-coding RNA expression, the use of just one of the stable reference genes produced a reliable measurement.