Interleukin-1 beta, tumor necrosis factor-alpha and lipopolysaccharide induce expression of monocyte chemoattractant protein-1 in calf aortic smooth muscle cells

J Tongji Med Univ. 2000;20(1):36-8. doi: 10.1007/BF02887671.

Abstract

To investigate whether interleukin-1 beta(IL-1 beta), tumor necrosis factor-alpha (TNF-alpha) and lipopolysaccharide (LPS) induce expression of monocyte chemoattractant protein-1 (MCP-1) mRNA and protein in calf aortic smooth muscle cells(SMCs), calf aortic SMCs were cultured by a substrate-attached explant method. The cultured SMCs were used between the third to the fifth passage. After the cells became confluent, the SMCs were exposed to 2 ng/ml IL-1 beta, 20 ng/ml TNF-1 alpha and 100 ng/ml LPS respectively, and the total RNA of SMCs which were incubated for 4 h at 37 degrees C were extracted from the cells by using guanidinium isothiocyanate method. The expression of MCP-1 mRNA in SMCs was detected by using dot blotting analysis using a probe of gamma-32P-end-labelled 35-mer oligonucleotide. After a 24-h incubation, the media conditioned by the cultured SMCs were collected. The MCP-1 protein content in the conditioned media was determined by using sandwich ELISA. The results were as follows: Dot blotting analysis showed that the cultured SMCs could express MCP-1 mRNA. After a 4-h exposure to IL-1 beta, TNF-alpha and LPS, the MCP-1 mRNA expression in SMCs was increased (3.6-fold, 2.3-fold and 1.6-fold, respectively). ELISA showed that the levels of MCP-1 protein in the conditioned media were also increased (2.9-fold, 1.7-fold and 1.1-fold, respectively). The results suggest that calf aortic SMCs could express MCP-1 mRNA and protein. IL-1 beta and TNF-alpha can induce strong expression of MCP-1 mRNA and protein, and the former is more effective than the latter.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aorta / metabolism
  • Cattle
  • Chemokine CCL2 / biosynthesis*
  • Chemokine CCL2 / genetics
  • Interleukin-1 / pharmacology*
  • Lipopolysaccharides / pharmacology*
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / metabolism*
  • RNA, Messenger / biosynthesis
  • Tumor Necrosis Factor-alpha / pharmacology*

Substances

  • Chemokine CCL2
  • Interleukin-1
  • Lipopolysaccharides
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha