Microvascular and macrovascular endothelial cells produce different constrictor substances

Marilyn K Glassberg Csete, K. B. Nolop, J. T. Jackowski, W. M. Abraham, Adam Wanner, U. S. Ryan

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The media from cultured microvascular and macrovascular endothelial cells (conditioned media, CM) were collected and tested for constrictor activity in sheep coronary artery rings and tracheal smooth muscle strips in vitro (isometric force), expressed as percentage of contraction produced by 80 mM KCl. Both microvascular (micro) and macrovascular (macro) CM caused a sustained slow-onset contraction (P < 0.05) of the coronary artery rings by 71 ± 10% (micro; n = 7) and 67 ± 8% (macro; n = 6) and tracheal smooth muscle strips by 33 ± 14% (micro; n = 6) and 34 ± 6% (macro; n = 11); the calcium antagonist gallopamil (10-7 M) attenuated these effects by 25-55%. Unconditioned medium and medium conditioned by cultured tracheal smooth muscle cells had no constrictor activity on coronary artery rings or tracheal smooth muscle strips. Synthetic endothelin (ET-1) also produced contraction of coronary artery rings and tracheal smooth muscle strips. The mean levels of ET-1 measured by radioimmunoassay were 1,200 pg/ml in the macro CM and 33 pg/ml in the micro CM. Depleting macro CM of ET-1 by affinity columns constructed with protein A agarose and anti-ET-1 antibody removed the contractile activity for coronary artery rings and tracheal smooth muscle strips. Thus ET-1 did not appear to be the contractile substance in the micro CM. Preliminary characterization of the contractile substance in micro CM revealed that it was heat stable, had a molecular weight of <10,000, was inactivated by trypsin, and retained its activity after two cycles of freeze- thawing. These observations suggest that 1) macrovascular and microvascular endothelial cells release different contractile substances that have a greater activity on sheep vascular than airway smooth muscle, 2) the contractile substance released from macrovascular endothelial cells is ET-1, and 3) the contractile responses involve calcium influx.

Original languageEnglish
Pages (from-to)1681-1686
Number of pages6
JournalJournal of Applied Physiology
Volume72
Issue number5
StatePublished - Jan 1 1992

Fingerprint

Conditioned Culture Medium
Endothelial Cells
Smooth Muscle
Coronary Vessels
Sheep
Gallopamil
Calcium
Staphylococcal Protein A
Endothelin-1
Vascular Smooth Muscle
Sepharose
Trypsin
Smooth Muscle Myocytes
Radioimmunoassay
Hot Temperature
Molecular Weight
Antibodies

Keywords

  • airway smooth muscle
  • contractile peptides
  • endothelin
  • endothelium
  • vascular smooth muscle

ASJC Scopus subject areas

  • Endocrinology
  • Physiology
  • Orthopedics and Sports Medicine
  • Physical Therapy, Sports Therapy and Rehabilitation

Cite this

Microvascular and macrovascular endothelial cells produce different constrictor substances. / Glassberg Csete, Marilyn K; Nolop, K. B.; Jackowski, J. T.; Abraham, W. M.; Wanner, Adam; Ryan, U. S.

In: Journal of Applied Physiology, Vol. 72, No. 5, 01.01.1992, p. 1681-1686.

Research output: Contribution to journalArticle

Glassberg Csete, Marilyn K ; Nolop, K. B. ; Jackowski, J. T. ; Abraham, W. M. ; Wanner, Adam ; Ryan, U. S. / Microvascular and macrovascular endothelial cells produce different constrictor substances. In: Journal of Applied Physiology. 1992 ; Vol. 72, No. 5. pp. 1681-1686.
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AB - The media from cultured microvascular and macrovascular endothelial cells (conditioned media, CM) were collected and tested for constrictor activity in sheep coronary artery rings and tracheal smooth muscle strips in vitro (isometric force), expressed as percentage of contraction produced by 80 mM KCl. Both microvascular (micro) and macrovascular (macro) CM caused a sustained slow-onset contraction (P < 0.05) of the coronary artery rings by 71 ± 10% (micro; n = 7) and 67 ± 8% (macro; n = 6) and tracheal smooth muscle strips by 33 ± 14% (micro; n = 6) and 34 ± 6% (macro; n = 11); the calcium antagonist gallopamil (10-7 M) attenuated these effects by 25-55%. Unconditioned medium and medium conditioned by cultured tracheal smooth muscle cells had no constrictor activity on coronary artery rings or tracheal smooth muscle strips. Synthetic endothelin (ET-1) also produced contraction of coronary artery rings and tracheal smooth muscle strips. The mean levels of ET-1 measured by radioimmunoassay were 1,200 pg/ml in the macro CM and 33 pg/ml in the micro CM. Depleting macro CM of ET-1 by affinity columns constructed with protein A agarose and anti-ET-1 antibody removed the contractile activity for coronary artery rings and tracheal smooth muscle strips. Thus ET-1 did not appear to be the contractile substance in the micro CM. Preliminary characterization of the contractile substance in micro CM revealed that it was heat stable, had a molecular weight of <10,000, was inactivated by trypsin, and retained its activity after two cycles of freeze- thawing. These observations suggest that 1) macrovascular and microvascular endothelial cells release different contractile substances that have a greater activity on sheep vascular than airway smooth muscle, 2) the contractile substance released from macrovascular endothelial cells is ET-1, and 3) the contractile responses involve calcium influx.

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