Interleukin 12 on extracellular matrix-related factors and functional characteristics of equine endometrial fibroblasts.

CONTEXT: Tissue fibrosis is described as the excessive accumulation of extracellular matrix (ECM) components. This process is associated with inflammation, and the interaction between fibroblasts and immune cells plays a key role, driving the progression of fibrosis. Results of recent study suggest that interleukin (IL)-12 may play a role in the processes associated with the development of fibrosis. This proinflammatory cytokine is synthesized and released by macrophages, dendritic cells, and B cells.

AIMS: The objective of this study was to determine: (i) the expression of IL-12 and its receptor in different mare endometrial categories; (ii) the effects of IL-12 on the expression of ECM-related factors; and (iii) endometrial fibroblast functional characteristics.

METHODS

The mRNA expression of IL-12 subunits and IL-12 receptor was determined using quantiative polymerase chain reaction (qPCR). The expression of ECM-related factors and functional characteristics in endometrial fibroblasts were determined using qPCR, Western blotting, zymography with bromodeoxyuridine and scratch assays.

KEY

RESULTS

IL-12Rβ2 mRNA expression was upregulated in category IIB endometrium during the mid-luteal phase compared with the follicular phase of the estrous cycle. The IL-12 treatment of endometrial fibroblasts increased mRNA expression of COL1A1, COL3A1, ACTA2, and LOXL2, as well as matrix metalloproteinase (MMP)-9 and MMP3, with elevated pro-MMP2 and MMP9 gelatinolytic activity. Moreover, IL-12 reduced fibroblast proliferation after 96 h, without effect on migration.

CONCLUSIONS

The findings indicate that IL-12 has a direct impact on the mRNA expression of fibrotic markers, MMP-2 and MMP-9 gelatinolytic activity and fibroblast proliferation.

IMPLICATIONS: This suggests a potential role for IL-12 in the processes associated with ECM remodeling.