研究目的
To determine whether carteolol eye drops, a b-adrenoceptor antagonist used as an intraocular hypotensive agent, has protective effects against the light-induced oxidative stress in retina.
研究成果
Carteolol application alleviated retinal damage from light exposure in vivo and photoreceptor cell death from oxidative stress in vitro, through mechanisms such as enhancing antioxidative potential by inducing TRX1 and GPX1 and decreasing intracellular ROS production, potentially via beta-adrenoceptors.
研究不足
The study did not find direct radical scavenging ability for carteolol, and the neuroprotective effects may involve indirect mechanisms. High doses were required in vitro, and intrinsic sympathetic activity differences between carteolol and timolol were noted but not fully explored.
1：Experimental Design and Method Selection:
The study used in vivo experiments on rats to assess retinal damage from light exposure and in vitro experiments on 661 W cells to evaluate oxidative stress. Methods included ERG recordings, histological examinations, immunostaining, RT-PCR, cell death assays, caspase activity measurements, ROS production measurements, DPPH radical scavenging assay, and TBARS assay.
2：Sample Selection and Data Sources:
Brown-Norway pigmented rats were used for in vivo studies, and 661 W murine photoreceptor cells for in vitro studies. Samples included retinal tissues and cell cultures.
3：List of Experimental Equipment and Materials:
Equipment included electroretinogram systems, microscopes, microplate readers, PCR machines, and spectrophotometers. Materials included carteolol hydrochloride, timolol maleate, antibodies, dyes, and culture media.
4：Experimental Procedures and Operational Workflow:
Rats were pre-treated with carteolol or saline, exposed to light, and assessed for ERG, ONL thickness, and 8-OHdG. Cells were treated with BSO/glutamate and assessed for cell death, caspase activity, and ROS. Assays for DPPH scavenging and TBARS were conducted.
5：Data Analysis Methods:
Statistical analyses used Student's t-tests and ANOVA with Tukey-Kramer or Dunnett's tests, performed with SPSS software.
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Toshiba FLR20S$W/M
FLR20S$W/M
Toshiba Co. Ltd.
White fluorescent light source for light exposure in in vivo studies.
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Fetal bovine serum
FBS
Thermo Fisher Scientific
Supplement for cell culture media.
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Olympus IX70
IX70
Olympus
Inverted epifluorescence microscope for examining and photographing cell specimens.
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Varioskan Flash 2.4
Varioskan Flash 2.4
Thermo Fisher Scientific
Microplate reader for measuring luminescence in caspase assays.
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Mikelan ophthalmic solution
2%
Otsuka Pharmaceutical Co., Ltd.
Used as the test sample for in vivo studies, containing carteolol hydrochloride.
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Midrin-P ophthalmic solution
Santen Pharmaceutical Co., Ltd.
Used for pupil dilation in rats before light exposure.
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LE-1000
LE-1000
Mayo Corp.
Bipolar contact lens electrode with embedded light emitting diode for ERG recordings.
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Vectastain Elite ABC kit
Vector Lab.
Used for immunohistochemical staining to visualize immunoreactivity.
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Trizol
Invitrogen, Life Technologies Inc.
Used for total RNA extraction from retinal tissues.
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SuperScript III First-Strand Synthesis System
Invitrogen
Used for cDNA synthesis from total RNA.
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Platinum Taq DNA polymerase mix
Invitrogen
Used for PCR amplification of cDNA.
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IS4000R
IS4000R
Eastman Kodak Company.
UV transilluminator/digital imaging system for photographing PCR products.
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Dulbecco's modified Eagle medium
DMEM
Nacalai Tesque Inc
Culture medium for maintaining 661 W cells.
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Hoechst 33342
Invitrogen
Fluorescent stain for dead and live cells in cell death assays.
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Propidium iodide
PI
Invitrogen
Fluorescent stain for dead cells in cell death assays.
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Caspase-Glo 3/7 Assay
Promega
Used for measuring caspase-3/7 activity in cells.
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CM-H2DCFDA
Thermo Scientific
Used for measuring intracellular reactive oxygen species production.
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