Inhibition of release of tumor necrosis factor-alpha from human vascular tissue and smooth muscle cells by glucocorticoids.
OBJECTIVE
Based on our previous study that bacterial lipopolysaccharide stimulates release of tumor necrosis factor (TNF)-alpha from human vascular tissue and smooth muscle cells, we tested the hypothesis that release of TNF could be inhibited by pretreatment with glucocorticoids.


METHODS
Prospective, repeated-measures analysis of concentration-response relationships.


METHODS
Academic anesthesiology research laboratory.


METHODS
Segments of internal mammary artery and saphenous vein were obtained during coronary artery bypass surgery.


METHODS
None.


RESULTS
Confluent human smooth muscle cells, cultured from saphenous vein and internal mammary artery, were exposed to 20 micrograms/mL of bacterial lipopolysaccharide following pretreatment for 18 hrs with either 0.1, 1.0, or 10.0 microM of dexamethasone. At 1, 3, 6, 18, and 24 hrs, the culture medium was removed and analyzed for biologically active TNF-alpha using the L929 cell cytotoxicity assay. Smooth muscle cells exposed to bacterial lipopolysaccharide but not treated with dexamethasone served as controls. In control internal mammary cells, bacterial lipopolysaccharide stimulated TNF-alpha release in a time-dependent manner to a peak of 36 +/- 2.3 U/mg of cell protein at 6 hrs, compared with 0.7 +/- 0.3 U/mg of cell protein in cells not exposed to lipopolysaccharide. Dexamethasone inhibited bacterial lipopolysaccharide-stimulated release at all time points in a concentration-dependent manner. For instance, at 6 hrs, TNF-alpha was 12 +/- 2.2, 6.9 +/- 1.7, and 2.3 +/- 0.9 U/mg of cell protein for cells pretreated with 0.1, 1.0, and 10.0 microM of dexamethasone, respectively (p < .05 vs. control). In separate experiments, segments of internal mammary artery and saphenous vein were obtained from five patients who received 1 g of methylprednisolone intravenously during induction of anesthesia, and from seven patients who did not receive methylprednisolone. Bacterial lipopolysaccharide induced release of TNF-alpha from vascular tissues of untreated patients in a time-dependent manner (e.g., 733 +/- 44 U/g of tissue at 6 hrs in saphenous vein). In contrast, in patients treated with methylprednisolone, bacterial lipopolysaccharide did not stimulate release from vascular tissues incubated for up to 24 hrs.


CONCLUSIONS
These results indicate that human vascular tissue, particularly the smooth muscle cell, may be a source of TNF-alpha and that glucocorticoids inhibit release stimulated by bacterial lipopolysaccharide.
Newman
W H
WH
Department of Anesthesiology, Mercer University School of Medicine, Macon, GA, USA.
Zhang
L M
LM
Leeper-Woodford
S K
SK
Shaker
I J
IJ
Erceg
S K
SK
Castresana
M R
MR
D002478:Cells, Cultured; D003907:Dexamethasone / Q000627:therapeutic use*; D004353:Drug Evaluation, Preclinical; D005938:Glucocorticoids / Q000627:therapeutic use*; D006801:Humans; D008070:Lipopolysaccharides; D008323:Mammary Arteries / Q000187:drug effects*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D012501:Saphenous Vein / Q000187:drug effects*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000187:drug effects* / Q000276:immunology
D005938:Glucocorticoids; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone