角膜结膜及泪器病             3107                                      PO0377

　　Proteomic Analysis of the Mice Cornea with Regard to Corneal Dystrophies

　　Ye Wang, Yiqiang Wang*, Xuemin Zhang

　　山东省眼科研究所 266071

　　To improve our understanding the pathogenesis of corneal dystrophy we utilized proteomic approach to study the protein expression profiles from the mice with hereditary corneal dystrophy.

　　Methods: Animals used in this experiment were of a inbreed strain of BABL/C mice that have spontaneously corneal dystrophies. At various age after birth of mice, corneas were harvested, solubalized, and analyzed by two-dimensional gel electrophoresis. Firstly, non-linear pH gradient strips (pH 3-10 NL) of 18 cm in length were rehydrated overnight with protein sample solutions on the isoelectric focusing flat bed. Then, strips were loaded onto 13% SDS-gel and run to resolve the protein samples on the second dimension. Interested protein spots were excised from the gel and digested with trypsin. Resultant peptides cocktails were applied to target plates for MALDI-TOF.

　　Results:  Gels were analyzed with ImageMaster TM 2D Platinum software. Spectra were used to search protein databases using Mascot. A Mascot score above 75 indicated a significant potential match between a database sequence and the identified sequences from mass spectrometry. Protein spots that changed greater than five-fold in level of expression ( relative to control ) were excised for identification. Six spots were successfully identified by MALDI/TOF. They included crystallin alpha A (19kD), crystallin beta A1 (25kD) and crystallin beta B2 (23kD). These proteins were all up-regulated.
 
　　Conclusion: In this study we looked at the proteomic analysis of the diseased mice corneas and found that crystallins were highly upregulated with the progression of corneal dystrophy in mice. The alpha-crystallins are the members of the small heat shock protein family and have chaperone-like activity preventing the aggregation of proteins. The beta-crystallins are related to microbial oxidative stress proteins and have been implicated in the development of cataracts. The abundant taxon-specific proteins in the lens and cornea are often similar or identical to metabolic enzymes and, therefore, termed enzyme-crystallins. The function of the corneal enzyme-crystallins is not clear, but it has been suggested that they serve catalytic as well as structural roles reminiscent to the alpha- and beta-crystallins in the lens and thereby contribute to the transparency and optical properties of the corneal cells. This study provided us inspiration and supporting evidence in understanding the molecular basis of corneal dystrophies, and further studies are underway.

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