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A. Higuchi, Y. Okubo, M. Watanabe, M. Hirashima, S. Shimmura, K. Tsubota; Identification of a Candidate for the Treatment of Dry Eye From Serum . Invest. Ophthalmol. Vis. Sci. 2006;47(13):252.
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Since autologous serum is useful for the treatment of severe dry eye, serum components may be a potential candidate for the treatment of dry eye. We tried to identify useful serum components for the treatment of dry eye.
Conjunctival epithelial cell line, CCL20.2 (CCL) requires 10% fetal bovine serum (FBS) in the medium to survive. When serum is removed from the culture medium, CCL undergo apoptosis. We choose the ratio of apoptosis of CCL as an index of purification. Human serum was fractionated by column chromatography and each fraction was added to the medium in exchange for FBS to assay the ratio of apoptosis of the cells. To confirm the effect of partial purified sample on the treatment of dry eye, the sample was applied to eye drops for the treatment of dry eye model rat. Since the treatment using partial purified sample was significantly effective compared with PBS treatment, we tried the further purification of this sample using by ion exchange column chromatography. SDS–PAGE of the purified fraction showed 2 of the major bands (band a and b). To identify the each band, each band was cut from the gel and applied on Edman method.
Protein b, which is originated band b, suppressed the ratio of apoptosis of CCL induced by serum deprivation. Experiment of eye drop of protein b using dry eye model rat showed that the fluorescein staining area of cornea treated by protein b was significantly small compared with PBS treatment. Furthermore, expression of MMP–2 and –9 in cornea decreased by treatment with protein b.
This study is the first trial of application of protein b on the treatment of dry eye. It is suggested that protein b is a candidate for the treatment of dry eye.
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