Mast cell tryptase stimulates the synthesis of type I collagen in human lung fibroblasts.

Jennifer Cairns, Andrew F. Walls

Journal of Clinical Investigation · 1997 · 320 citations · 56 references

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TL;DR

Mast cell activation is a hallmark of chronic inflammation that can drive fibrosis through increased fibroblast collagen production. The study aimed to determine whether mast‑cell tryptase can stimulate collagen synthesis in human lung fibroblasts (MRC‑5). Tryptase was purified from human lung tissue by ion‑exchange and affinity chromatography and then applied to MRC‑5 cells to assess its effects. Tryptase at 18–36 mU/ml enhanced cell proliferation, DNA synthesis (≈5×), collagen synthesis (≈2.5×), and collagenolytic activity (≈2×), effects largely dependent on its catalytic activity and producing predominantly type I collagen, suggesting that mast‑cell tryptase may promote fibrotic remodeling in inflammatory disease.

Abstract

Mast cell activation is a characteristic feature of chronic inflammation, a condition that may lead to fibrosis as a result of increased collagen synthesis by fibroblasts. We have investigated the potential of tryptase, the major protease of human mast cells, to stimulate collagen synthesis in the human lung fibroblast cell line MRC-5. Tryptase was isolated from human lung tissue by ion-exchange and affinity chromatography. At concentrations of 18 and 36 mU/ml, tryptase stimulated both an increase in cell numbers, and a fivefold increase in DNA synthesis as determined by methyl-[3H]thymidine incorporation. Similar concentrations of tryptase resulted in a 2.5-fold increase in collagen synthesis as determined both by incorporation of [3H]proline into collagen, and by assay of hydroxyproline concentrations in the supernatants. There was also a twofold increase in collagenolytic activity in the culture medium after tryptase treatment, indicating that the increase in collagen synthesis was not a consequence of decreased collagenase production. All of these actions of tryptase were reduced in the presence of the protease inhibitors leupeptin and benzamidine hydrochloride, indicating a requirement for an active catalytic site. SDS-PAGE and autoradiographic analysis of the [3H]collagen produced by the cells revealed it to be predominantly type I collagen. Our findings suggest that the release of tryptase from activated mast cells may provide a signal for abnormal fibrosis in inflammatory disease.

References

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