IL-1-BETA AND IL-6 INDUCE HYPERPLASIA AND HYPERTROPHY OF CULTURED GUINEA-PIG AIRWAY SMOOTH-MUSCLE CELLS

Guinea pig airway smooth muscle (ASM) cells were maintained in a primary tissue culture (passages 1-3). Cells were exposed to human recombinant interleukin-1 beta (IL-1 beta; 20-100 pg/ml) or interleukin-6 (IL-6; 1-4 ng/ml) in the presence of indomethacin (1 mu g/ml) for up to 5 days. Proliferation of ASM cells was assessed with two techniques, direct counting of cells with a hemacytometer and [H-3]thymidine incorporation corrected for total protein content. Hypertrophy of ASM cells was assessed by [H-3]leucine incorporation (evaluation of protein synthesis), determination of total DNA content, DNA content per cell, and protein content per cell. We observed that the exposure of ASM cells to human recombinant IL-1 beta or IL-6, in all studied concentrations, significantly increased the number of cells as well as [H-3]thymidine incorporation into ASM cells. We also found that exposure of ASM to these two cytokines increased [H-3]leucine incorporation into the ASM cells and increased protein content and DNA content per single cell. These changes were also concentration dependent. We conclude that the two proinflammatory cytokines, IL-1 beta and IL-6, which are present in asthmatic lungs, increased the proliferation of ASM cells (hyperplasia) as well as their overall size and size of their nuclei, as measured by biochemical markers. These findings are compatible with the presence of ASM hypertrophy.