研究目的
To find out the effects of nuclear factor erythroid 2-related factor/heme oxygenase-1 (Nrf2/HO-1) pathway on retinal ganglion cell (RGC) injury in glaucoma.
研究成果
Activation of the Nrf2/HO-1 pathway significantly reduces apoptosis and injury of retinal ganglion cells in rats with chronic ocular hypertension, offering a protective effect in glaucoma and suggesting a potential clinical target for preventing RGC degeneration.
研究不足
The study is limited to a rat model, which may not fully replicate human glaucoma conditions. Potential areas for optimization include the use of more advanced imaging techniques or larger sample sizes to enhance reliability.
1：Experimental Design and Method Selection:
The study involved constructing chronic ocular hypertension (COH) rat models of glaucoma to investigate the effects of the Nrf2/HO-1 pathway on RGC injury. Methods included TUNEL staining for apoptosis detection, qRT-PCR and western blotting for gene and protein expression analysis, and HE staining for morphological observation.
2：Sample Selection and Data Sources:
A total of 40 male SD rats weighing 200–300 g were used for model construction, and another 24 SD rats were divided into groups (normal, model, model + tBHQ) for treatment studies. Data on IOP and RGC numbers were collected at specified time points.
3：List of Experimental Equipment and Materials:
Equipment included an ophthalmic operating microscope (OPMI VISU 140, Carl Zeiss), portable tonometer TonoLab (Mento O&O), microscope (Leica), and various kits and reagents such as TUNEL mixture (Promega), antibodies (Abcam), and tBHQ (Sigma-Aldrich). Materials included sutures, anesthetics, and staining agents.
4：Experimental Procedures and Operational Workflow:
Rats were anesthetized, COH models were created by ligating veins in the eye, IOP was measured at intervals, and treatments (tBHQ or vehicle) were administered. After euthanasia, retinal tissues were processed for FG tracing, HE staining, TUNEL staining, qRT-PCR, and western blotting.
5：Data Analysis Methods:
Data were analyzed using SPSS 22.0 software with Student's t-test for two-group comparisons and one-way ANOVA for multiple groups, with P < 0.05 considered significant.
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ophthalmic operating microscope
OPMI VISU 140
Carl Zeiss
Used for surgical procedures to construct the chronic ocular hypertension model in rats.
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portable tonometer
TonoLab
Mento O&O
Used to detect intraocular pressure (IOP) in rat eyes.
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microscope
Leica
Used for observing pathological changes in retinal tissues.
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TUNEL mixture
Promega
Used for detecting apoptosis in retinal ganglion cells via TUNEL staining.
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antibodies
ab137550, ab13248, ab8245
Abcam
Used in western blotting to detect expressions of Nrf2, HO-1, and GAPDH proteins.
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tBHQ
Sigma-Aldrich
Used as an Nrf2 activator in oral gavage to treat rats and study its effects on the Nrf2/HO-1 pathway.
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