Effect of LDL derived from ferroHb- and activated PMN–containing plasma on HO-1 and ferritin in endothelial cells.
LDL derived from ferroHb- and activated PMN–containing plasma induces HO-1 and ferritin in endothelial cells. (A) For HO-1 mRNA analysis, endothelium was treated with native LDL (first lane); LDL isolated from plasma containing 20 μM ferroHb (second lane), 20 μM ferroHb and PMA (500 ng/mL)–activated PMNs (107/mL) (third lane), PMA-activated PMNs alone (fourth lane), or 20 μM ferroHb and PMA (fifth lane); or LDL from plasma containing 20 μM ferroHb and resting PMNs (sixth lane) for 60 minutes in medium 199 followed by a 4-hour incubation in medium alone. Total cell RNA was isolated, and Northern blots of endothelial cell RNA were probed with biotin-labeled HO-1 cDNA probe. (B) Autoradiograph was quantified by video densitometry. (C) The corresponding 28S and 18S ribosomal RNA of the Northern blot in panel A. (D) HO enzyme activity measured at 8 hours. (E) Ferritin content was measured for the same groups as in panel A. Results represent mean ± SE of at least 3 experiments performed in duplicate.
