Figure 2

Dysregulated eicosanoid production and expression of enzymes and proteins responsible for eicosanoid generation from young AMs compared to adult AMs. AMs were harvested from adult (~10 weeks old) and young (day 7 and 14) rats and allowed to adhere overnight. A. and B. Supernatants were collected from AM cultures that were either unstimulated (A) or stimulated with LPS (B, 500 ng) for 24 h. The amount of PGE₂ was quantified by specific EIA. C. and D. Supernatants were collected from AM cultures either with media for 24 h (C) or with Ca²⁺ ionophore A23187 (D, 5 μM) for 1 h. The amount of LTB₄ was quantified by specific EIA. E. and F. RNA was extracted from the cells, converted into cDNA and the relative expression was determined using semi-quantitative real-time PCR. The primers for specific genes of interest are located in Table 1. For each sample, the expression of the gene of interest was normalized to β-actin and the average of the adult AMs was set to 1. For EIA, data shown are representative of three independent experiments preformed at least in triplicate. For real time PCR experiments, data represents the mean ± SEM of pooled samples from 3 independent experiments. (*p < 0.05, **p < 0.01, and p < 0.001 when compared to adult AMs).