Abstract

P 145

The role of relaxin2 and relaxin-like factor at the ocular surface

Ulrike Hampel1, Thomas Klonisch2, Friedrich Paulsen1
1Institut für Anatomie und Zellbiologie, Martin-Luther-Universität Halle-Wittenberg, Halle/Saale, 2Department of Human Anatomy and Cell Science, University of Manitoba, Winnipeg, Manitoba, Kanada

Objective
The pregnancy related hormones relaxin 2 (RLN2) and relaxin-like factor (INSL3) are predominantly known for their effects on the reproductive system. They induce remodelling of the extracellular matrix (ECM) by influencing matrix metalloproteinases (MMPs) and inhibitors of matrix metalloproteinases (TIMPs). Dry eye syndrome is associated with ECM remodelling and chronic inflammation of the ocular surface and is accompanied by activation of MMPs. In recent investigations we detected human RLN2, INSL3 and their cognate relaxin-like receptors LGR7 and LGR8 in human corneal (Araki-Sasaki, HCE), conjunctival (IOBA-NHC, HCjE) and sebocyte (SZ95, SC) cell lines. In this study, we investigated potential effects of human RLN2 and INSL3 on cell proliferation and migration as well as MMP and TIMP expression in HCE, HCjE, and SC.
Methods
Proliferation was tested by an assay in cultured HCE, HCjE, and SC after stimulation with different concentrations of RLN2 and INSL3. A scratch assay was established to investigate the migration of HCE and HCjE cells after 24h stimulation with RLN2 and INSL3. Real-time PCR analysis was used to detect the influence of RLN2 and INSL3 on MMP2, MMP9, MMP13, TIMP1, and TIMP2 after 24h stimulation.
Results
Stimulation of HCE, HCjE, and SC with RLN2 and INSL3 significantly increased cell proliferation in all three cell types. No effect was achieved in primary corneal fibroblasts that were used for comparison. RLN2 and INSL3 increase migration of HCE and HCjE cells after 24h stimulation significantly. After 24h stimulation of HCE, HCjE and SC with RLN2 and INSL3 the mRNA expression levels of MMP2 increased. In HCE and SC the mRNA concentration of TIMP1 was up-regulated after stimulation, whereas it was downregulated in HCjE. TIMP2 mRNA expression levels were elevated in HCjE and SC, but were not affected in HCE. mRNA concentrations of MMP9 and MMP13 were not significantly influenced by RLN2 or INSL3 stimulation in all three cell lines.
Conclusions
Taken together, our results show that RLN2 and INSL3 enhance cell proliferation and migration. Furthermore both hormones influence MMP2, TIMP1 and TIMP2 expression in HCE, HCjE and SC. It remains to be clarified whether RLN2 and INSL3 may have a similar role in vivo.

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