研究目的
To evaluate the neuroprotective effect of melatonin on retinal ganglion cells subject to oxidative stress using a model of oxidative glutamate toxicity in combination with BSO.
研究成果
Melatonin demonstrated significant neuroprotective and antiapoptotic effects in RGCs subjected to oxidative stress induced by BSO-GLUT, both in vitro and in vivo. It increased cell survival, reduced apoptosis, and minimized ultrastructural damage. These findings suggest melatonin is a promising therapeutic agent for treating ocular neurodegenerative diseases like glaucoma when administered locally.
研究不足
The study used animal models (rabbits and chicken embryos), which may not fully replicate human conditions. The oxidative stress model with BSO-GLUT might not cover all aspects of human ocular neurodegenerative diseases. The duration of the study was limited to 9 days, and long-term effects were not assessed. ERG results showed no functional changes, indicating potential limitations in detecting early functional impairments.
1：Experimental Design and Method Selection:
The study used in vitro and in vivo models to assess the neuroprotective efficacy of melatonin against oxidative stress induced by BSO and glutamate in retinal ganglion cells. Methods included cell culture techniques, cytotoxicity assays, histological studies, immunohistochemistry (TUNEL assay), electroretinography (ERG), and transmission electron microscopy (TEM).
2：Sample Selection and Data Sources:
RGCs were purified from 8-day-old chicken embryos using an immunopanning method. In vivo experiments used New Zealand albino rabbits. Samples included control groups, BSO-GLUT treated groups, and BSO-GLUT + MEL treated groups.
3：List of Experimental Equipment and Materials:
Chemicals such as glutamate, BSO, melatonin, proteinase K, hydrogen peroxide, methyl green, glutaraldehyde, formaldehyde from Sigma-Aldrich; ApopTag Plus Peroxidase In situ Apoptosis Detection Kit from Chemicon International Invitrogen; anesthetics and antiseptics from various brands; DMEM medium for cell culture; syringes with 29-gauge needles; light microscope (Olympus BX41); camera (Infinity 1); image analysis software (Image J); ERG equipment (AKONIC BIO-PC); transmission electron microscope (Zeiss LEO 906E); ultramicrotome (JEOL JUM-7).
4：7).
Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: For in vitro studies, RGC cultures were treated with BSO, GLUT, and MEL for 24 and 48 hours, followed by cell survival assays (crystal violet technique) and TUNEL assays. For in vivo studies, rabbits received intravitreal injections of BSO-GLUT or BSO-GLUT + MEL, and after 9 days, retinas were analyzed histologically, with TUNEL, ERG, and TEM. ERG was performed at specific time points with standardized light stimuli.
5：Data Analysis Methods:
Data were analyzed using statistical methods such as student t-test and analysis of variance, with p < 0.01 considered significant. Image analysis was done with Image J software.
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