研究目的
Investigating the neuroprotective effects of adrenergic receptor (AR)-targeting pharmaceuticals, the α1-AR antagonist doxazosin or the α2-AR agonist guanabenz, against photoreceptor cell death in retinal detachment (RD).
研究成果
Pharmacological targeting of α1-AR with doxazosin or α2-AR with guanabenz effectively protects photoreceptors from apoptosis and preserves retinal function in a rat model of retinal detachment by inhibiting ROS overproduction and proinflammatory cytokine upregulation. These findings suggest adrenergic receptors as novel therapeutic targets for retinal diseases, with potential for further exploration of FDA-approved drugs like doxazosin and guanabenz.
研究不足
The study did not establish the precise localization of NADPH oxidase (NOX) in the retina, did not investigate mitochondria-derived ROS involvement, did not assess intraocular concentrations of drugs, and used a rat model which may not fully translate to humans. High-dose and long-term treatment with guanabenz caused side effects like drowsiness and hypoactivity.
1：Experimental Design and Method Selection:
The study used an experimental RD model in Brown-Norway rats induced by subretinal injection of sodium hyaluronate. Pharmacological interventions with doxazosin (α1-AR antagonist) and guanabenz (α2-AR agonist) were administered systemically. Methods included ELISA for oxidative stress and cytokine quantification, immunofluorescence, Western blotting, TUNEL assay, histological examination, and electroretinography (ERG).
2：Sample Selection and Data Sources:
Adult male Brown-Norway rats (8-10 weeks; 180-250 g) were used. Retinal tissues and subretinal fluid samples were collected at various time points post-RD induction.
3：List of Experimental Equipment and Materials:
Sodium hyaluronate (10 mg/ml, LG Life Sciences), doxazosin (Selleckchem Cat# S1324), guanabenz (Selleckchem Cat# S4065), apocynin (Selleckchem Cat# S2425), ELISA kits (R&D Systems), DHE probe (Molecular Probes), TUNEL kit (Roche), antibodies (e.g., anti-GFAP, anti-COX-2), confocal microscope (Leica TCS SP8), ERG system (Phoenix Research Laboratories Ganzfeld).
4：Experimental Procedures and Operational Workflow:
Rats were anesthetized, RD was induced via subretinal injection, drugs were administered intraperitoneally daily starting 1 hour before RD. Tissues were harvested at specified times for analysis. ROS detection used DHE staining, apoptosis was assessed by TUNEL, histology by H&E staining, protein expression by Western blot, and retinal function by ERG.
5：Data Analysis Methods:
Statistical analyses were performed using IBM SPSS Statistics (version 23) with Student's t-test or one-way ANOVA. Data are expressed as mean ± standard deviation, with P < 0.05 considered significant.
独家科研数据包，助您复现前沿成果，加速创新突破
获取完整内容-
confocal microscope
TCS SP8
Leica
Used for immunofluorescence imaging of retinal sections.
-
sodium hyaluronate
10 mg/ml
LG Life Sciences
Used for subretinal injection to induce retinal detachment in rats.
-
doxazosin
S1324
Selleckchem
α1-AR antagonist administered intraperitoneally to inhibit adrenergic receptor signaling.
-
guanabenz
S4065
Selleckchem
α2-AR agonist administered intraperitoneally to activate adrenergic receptor signaling.
-
apocynin
S2425
Selleckchem
NOX inhibitor administered intraperitoneally to inhibit NADPH oxidase.
-
ELISA kit
RLB00, MJE00, RTA00
R&D Systems
Used to quantify levels of IL-1β, MCP-1, and TNF-α in retinal lysates.
-
protein carbonyl content assay kit
STA-310
Cell Biolabs
Used to measure protein carbonyl content as a biomarker for oxidative stress.
-
dihydroethidium
D11347
Molecular Probes
ROS probe administered to detect superoxide anions in vivo.
-
TUNEL assay kit
11684795910
Roche Diagnostics
Used to detect apoptotic cells in retinal sections.
-
electroretinography system
Ganzfeld
Phoenix Research Laboratories
Used to record scotopic ERG responses to assess retinal function.
-
antibody
BA0738
Boster Biological Technology
Primary antibody against COX-2 for Western blotting.
-
antibody
60004-1-Ig
Proteintech Group
Primary antibody against GAPDH for Western blotting.
-
secondary antibody
SA00001-1, SA00001-2
Proteintech Group
HRP-labelled secondary antibodies for Western blotting.
-
imaging system
Amersham Imager 600
GE Healthcare
Used to detect chemiluminescence in Western blotting.
-
software
ImageJ 1.48v
National Institutes of Health
Used for image analysis and quantification of DHE-positive cells and TUNEL assays.
-
statistical software
SPSS Statistics version 23
IBM
Used for statistical analysis of experimental data.
-
登录查看剩余14件设备及参数对照表
查看全部
