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E Adachi-Usami, A Yokoyama, A Mizota, X Mo, H Negishi, T Oshitari, A Uemura; Electrically Applied Hsp27 Protected Retinal Ganglion Cells Against Ischemia-Reperfusion in Rat Retina . Invest. Ophthalmol. Vis. Sci. 2002;43(13):771.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To determine whether the heat shock protein 27 (Hsp27) can rescue retinal ganglion cells (RGCs) of rats from ischemia-reperfusion injury. Methods: Nine-week-old Male Wistar (200-250g) rats were used in the present experiment. Under the anesthesia with urethane, ketamine and xylazine, retinal ischemia was induced by clamping the ophthalmic artery within the dural sheath of the optic nerve. Immediately after removing the clamp and beginning the reperfusion, 5µ of Hsp27 protein solution, containing 6.5 µ g protein, was injected into the vitreous and electroporation was applied. To determine whether Hsp27 entered the RGCs, anti-Hsp27 immunohistochemistry was performed. The retinal damage was evaluated by counting the number of RGCs retrogradely labeled by 1,1'-dioctadecyl-3, 3,3', 3'-tetramethylindocarbocyanine percholorate (diI) injected into the superior colliculus, and also by comparing the ratio of TUNEL-positive to all RGCs in the RGC layer. Results: By immunohistochemical study, electroporation successfully delivered Hsp27 protein into RGCs. In the Hsp27 electroinjected group, the number of RGCs 7 days after ischemia-reperfusion was significantly greater than in the control groups. The ratio of TUNEL-positive to all RGCs was lower in the group electroinjected with Hsp27 protein. Conclusion: With electroporation Hsp27 can enter into RGCs. Electroporation of Hsp27 protein into RGCs increased the resistance of the RGCs to apoptosis induced by ischemia-reperfusion injury.
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