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(Investigative Ophthalmology and Visual Science. 2000;41:1962-1970.)
© 2000 by The Association for Research in Vision and Ophthalmology, Inc.

Adrenomedullin in Cultured Human Retinal Pigment Epithelial Cells

Tetsuo Udono1,2, Kazuhiro Takahashi1, Masaharu Nakayama1, Osamu Murakami3, Yusuf K. Durlu2, Makoto Tamai2 and Shigeki Shibahara1

1 From the Department of Molecular Biology and Applied Physiology, 2 Department of Ophthalmology, and 3 Second Department of Internal Medicine, Tohoku University School of Medicine, Miyagi, Japan.

PURPOSE. To determine whether adrenomedullin (ADM), a vasorelaxant peptide is produced and secreted by human retinal pigment epithelial (RPE) cells, whether ADM expression is regulated by inflammatory cytokines and a growth factor, and whether ADM has proliferative effects on these cells.

METHODS. Production and secretion of ADM by cultured human RPE cells were examined by Northern blot analysis and radioimmunoassay. Regulation of the ADM expression by basic fibroblast growth factor, interferon (IFN)-{gamma}, tumor necrosis factor-{alpha}, interleukin (IL)-1ß, or all-trans-retinoic acid was studied. In addition, proliferative effects of ADM on human RPE cells were examined by modified 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.

RESULTS. ADM mRNA was expressed constitutively in all three human RPE cell lines (F-0202, D407, and ARPE-19) examined. Immunoreactive ADM was detected in the cultured media by radioimmunoassay. Sephadex G-50 column chromatography of the cultured medium showed a single peak eluting in the position of ADM-(1-52). Treatment with IFN-{gamma} or IL-1ß increased ADM mRNA levels and immunoreactive-ADM levels in the medium in dose- and time-dependent manners in ARPE-19 cells. Exogenously added ADM increased the number of F-0202 cells and ARPE-19 cells, and the treatment with ADM antibody or ADM-(22-52) (an ADM antagonist) decreased it.

CONCLUSIONS. Human RPE cells produced and secreted ADM. IFN-{gamma} and IL-1ß induced ADM expression in ARPE-19 cells. Furthermore, ADM stimulated proliferation of RPE cells. These results raise the possibility that ADM is related to the pathophysiology of some inflammatory and proliferative ocular diseases.




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