The aim of this study was to examine the effect of molecular hydrogen (H2) on the healing of alkali-injured cornea. The effects of the solution of H2 in phosphate buffered saline (PBS) or PBS alone topically applied on the alkali-injured rabbit cornea with 0.25 M NaOH were investigated using immunohistochemical and biochemical methods. Central corneal thickness taken as an index of corneal hydration was measured with an ultrasonic pachymeter. Results show that irrigation of the damaged eyes with H2 solution immediately after the injury and then within next five days renewed corneal transparency lost after the injury and reduced corneal hydration increased after the injury to physiological levels within ten days after the injury. In contrast, in injured corneas treated with PBS, the transparency of damaged corneas remained lost and corneal hydration elevated. Later results-on day 20 after the injury-showed that in alkali-injured corneas treated with H2 solution the expression of proinflammatory cytokines, peroxynitrite, detected by nitrotyrosine residues (NT), and malondialdehyde (MDA) expressions were very low or absent compared to PBS treated injured corneas, where NT and MDA expressions were present. In conclusion, H2 solution favorably influenced corneal healing after alkali injury via suppression of oxidative stress.

• MeSH
• aktiny metabolismus   MeSH
• cytokiny metabolismus   MeSH
• exprese genu účinky léků   MeSH
• hydroxid sodný toxicita   MeSH
• imunohistochemie MeSH
• interleukin-1beta genetika   metabolismus   MeSH
• keratin-12 metabolismus   MeSH
• keratin-3 metabolismus   MeSH
• králíci MeSH
• kyselina peroxydusitá metabolismus   MeSH
• malondialdehyd metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• oxidační stres účinky léků   MeSH
• poranění rohovky etiologie   metabolismus   patologie   MeSH
• rohovka metabolismus   patologie   MeSH
• vaskulární endoteliální růstový faktor A genetika   metabolismus   MeSH
• vodík farmakologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

PURPOSE: The purpose of this study was to investigate the expression of cytokeratin (CK) 8 in the corneoconjunctival epithelium. METHODS: In 17 cadaveric corneoscleral discs and 3 other discs, the presence of CK8 alone or CK8, together with CK3, CK15, vimentin, and integrin α6, was investigated by using indirect immunohistochemistry on radial cryosections. Four corneoscleral discs stored in organ culture were used for the preparation of tangential sections of the limbus and for the isolation of limbal epithelial cells and their subsequent cultivation. CK8 expression was examined by RT-PCR in the corneal, limbal, and conjunctival epithelium. RESULTS: Sixty percent of the cadaveric corneoscleral samples and all samples stored in organ culture revealed positivity for CK8 in the basal epithelial layer of the limbus. Positive basal cells formed a single line or separated clusters. The signal for CK8 became weaker toward the surface of the limbal epithelium. The colocalization of CK8 with vimentin and CK15 in the limbus was also found. CK3 showed only occasional positivity in some of the surface limbal cells. The expression of integrin α6 in the basal membrane was absent or decreased under the CK8-positive clusters. Cell cultures revealed strong positivity for CK8 in approximately 80% of the cultured cells, and CK8 expression in the cornea, limbus, and conjunctiva was determined by RT-PCR. CONCLUSIONS: The study demonstrates the strong expression of CK8 in limbal epithelial basal cells, which is maintained during the differentiation and migration of the limbal cells toward the central corneal epithelium.

• MeSH
• buněčné kultury MeSH
• dospělí MeSH
• epitelové buňky metabolismus   MeSH
• exprese genu MeSH
• fluorescenční mikroskopie MeSH
• fluorescenční protilátková technika nepřímá MeSH
• integrin alfa6 metabolismus   MeSH
• keratin-15 metabolismus   MeSH
• keratin-3 metabolismus   MeSH
• keratin-8 genetika   metabolismus   MeSH
• konjunktiva cytologie   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• limbus corneae cytologie   metabolismus   MeSH
• messenger RNA genetika   MeSH
• mladiství MeSH
• orgánové kultury - kultivační techniky MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• rohovkový epitel cytologie   metabolismus   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• vimentin metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH