ABSTRACT
Transplantation of ex vivo cultured limbal epithelial cells is proven effective in restoring limbal stem cell deficiency. The present study aimed to investigate the promoting effect of Y-27632 on limbal epithelial cell proliferation. Limbal explants isolated from human donor eyes were expanded three weeks on culture dishes and outgrowth of epithelial cells was subsequently subcultured for in vitro experiments. In the presence of Y-27632, the ex vivo limbal outgrowth was accelerated, particularly the cells with epithelial cell-like morphology. Y-27632 dose-dependently promoted the proliferation of in vitro cultured human limbal epithelial cells as examined by phase contrast microscopy and luminescent cell-viability assay 30 hours after the treatment. The colony forming efficacy determined 7 days after the treatment was enhanced by Y-27632 also in a dose-dependent manner. The number of p63- or Ki67-positive cells was dose-dependently increased in Y-27632-treated cultures as detected by immunofluorescent staining and western blotanalysis. Cell cycle analysis by flow cytometric method revealed an increase in S-phase proliferating cells. The epithelial woundclosure rate was shown to be faster in experimental group received topical treatment withY-27632 than the sham control using a rat corneal wounding model. These resultsdemonstrate that Y-27632 can promote both the ex vivo and in vitro proliferation oflimbal epithelial cell proliferation. The in vivo enhanced epithelial wound healingfurther implies that the Y-27632 may act as a new strategy for treating limbal stem cell deficiency.
Subject(s)
Amides/pharmacology , Epithelial Cells/pathology , Limbus Corneae/pathology , Protein Kinase Inhibitors/pharmacology , Pyridines/pharmacology , Wound Healing/drug effects , rho-Associated Kinases/antagonists & inhibitors , Animals , Apoptosis/drug effects , Biomarkers/metabolism , Cell Cycle/drug effects , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Colony-Forming Units Assay , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Humans , Keratin-12/metabolism , Ki-67 Antigen/metabolism , Rats, Sprague-Dawley , Tumor Suppressor Protein p53/metabolism , rho-Associated Kinases/metabolismABSTRACT
BACKGROUND: Abnormality in diffusion-weighted magnetic resonance imaging representing early changes of acute ischemic lesions in human and animal models of focal status epilepticus has been reported to correlate with clinical outcome. CASE REPORT: We reported a 35 year-old woman with initial status epilepticus, probably related to previous head injury with traumatic intracerebral hemorrhage. The presenting MRI showed reversible hyperintensity lesions on DWI, which is probably corresponding to the epileptogenic lesion. Similar abnormalities in the splenium as a remote effect were demonstrated in this case. CONCLUSION: The atrophic changes in the splenium and right parietal lobe in the follow-up MRI scans were supposed to correlate with the following neurological sequelae.
