Expression of Plasminogen Activator Inhibitor-1 (PAI-1) mRNA in Human Ciliary Processes and Determination of PAI-1 in Human Aqueous Humor
Meyer M. W.1, Depka M.2, Wilhelm C.2, Schroeder A.1, Czwalinna A.2, Erb C.1
1Augenklinik und 2Hämatologie der Medizinischen Hochschule Hannover
Purpose: The present study was performed to detect the gene expression encoding for PAI-1 in human ciliary processes, to determine the quantity of PAI-1 and to measure PAI-1 activity in human aqueous humor and cell culture supernatants.
Method: Total mRNA was extracted from human ciliary processes, which were cut off from freshly enucleated human eyes (n=8). Reverse transcribed PAI-1 mRNA was measured by real time PCR (TaqMan® PCR). In addition, we determined the concentration of PAI-1 antigen (Ag) in supernatants of human ciliary processes in culture (n=8) and in aqueous humor of patients undergoing cataract surgery (n=18) by using an ELISA (Zymutest® PAI-1 Ag). The activity of PAI-1 in human aqueous humor (n=30) was measured by using a specific chromogenic test (Coatest® PAI-1).
Results: PAI-1 mRNA was localized in human ciliary processes. Depending on the time interval of cultivation the PAI-1 Ag levels ranged from 43.1 to 152.3 ng/ml. The quantity of PAI-1 Ag in human aqueous humor ranged from 0 to 4.56 ng/ml (mean 0.46 ng/ml). In contrast, PAI-1 activity was detected in all samples of human aqueous humor. The activity ranged from 6.5 to 19.0 AU/ml.
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