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C Esser, JM Esser, G Welsandt, A Hueber, P Esser, N Kociok, GK Krieglstein, H Mietz; Detection and Regulation of MMP and TIMP in Tenon´s Fibroblasts . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3333.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Latanoprost reduces the amount of extracellular matrix production of the conjunctiva, which is performed by Tenon´s capsule fibroblasts. This effect may be controlled by metalloproteinases (MMP) and their natural inhibitors, the tissue inhibitors of metalloproteinases (TIMP). For this reason, we investigated the presence of MMP and TIMP in human Tenon´s capsule fibroblasts in more detail and the effect of latanoprost on the levels of these enzymes. Methods: Human Tenon´s fibroblasts were obtained during surgery and prepared for cell culture. Fourth passage cells were used. First, immunohistochemistry was performed. Then, RNA extraction and RT-PCR was done. In addition, an immunoblot analysis and flow cytometry were carried out on cells treated with latanoprost. Finally, a Real-time PCR was done to evaluate the presence of different forms of MMP and TIMP, which may be related to tissue remodeling. Results: Immunohistochemistry clearly showed the presence of MMP-3 and TIMP-2 in the cells. Controls were stained with beta actin. Similarly, the RNA extraction and RT-PCR showed the presence of the RNA in the fibroblasts. Using the immunoblot analysis, similar results were obtained. With the flow cytometry technique, an up-regulation of the signal from 0.97 in control cells up to 1.20 in latanprost-treated cells was measured for TIMP-2, while for MMP-3, the controls were 1.04 and the signal increased to 1.24 following latanoprost-treatment. With the specific Real-time PCR technique, the presence of the MMPs 1, 2, 3, 7, 9 and 14 was confirmed, as well for TIMPs 1 and 2. Conclusion: The MMPs and TIMPs detected in our experiments may play an important role in extracellular matrix remodeling. While the presence of MMP-3 and TIMP-2 was confirmed in various different assays, the confirmation of the presence of a larger variety of MMPs and TIMPs may be important for further strategies to interfere with peribulbar wound healing and to achieve a decrease of extracelular matrix accumulation.
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