Figure 4.

IL-1β–induced changes in brain-derived neurotrophic factor (BDNF) were mediated through the IL-1 receptor and occurred, in part, at the level of mRNA expression (real-time quantitative PCR). Eutopic endometriosis stromal cells (EESCs) were preincubated with 1 μg/mL IL-1ra for 1 hour before stimulation with 10 ng/mL IL-1β. A: IL-1ra neutralized the stimulation of BDNF by IL-1β. When the gels were over-run, resolution of the 32- and 14-kDa bands into doublets was noted. All three major isoforms were affected. β-Actin levels were not altered. Similar findings were noted in independent EESC preparations. B: Triplicate EESC cultures were incubated for 24 hours in the presence of 0 to 160 ng/mL recombinant IL-1β. Total RNA extracted from the cultures was reverse transcribed and subjected to quantitative PCR amplification. The BDNF data were normalized to glyceraldehyde-3-phosphate dehydrogenase mRNA levels, using the formula 2^ΔΔct. Similar findings were noted in independent EESC preparations. n = 3 independent EESC preparations (A and B). ^∗P < 0.05 versus control (analysis of variance with Scheffé’s post hoc test).