{"doi":"10.1007/s10753-024-02152-w","title":"Triamcinolone Acetonide Protects Against Light-Induced Retinal Degeneration by Activating Anti-Inflammatory STAT6/Arg1 Signaling in Microglia","abstract":"All procedures conformed to the Association for Research in Vision and Ophthalmology (ARVO) Statement for the Use of Animals in Ophthalmic and Vision Research and were approved by the Animal Use and Care Committee of Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou, China. Two hundred specific pathogen-free (SPF) BALB/c mice (6–8 weeks old, weighing 19–25 g) were obtained from Ruisiyuan Biotech (Zhaoqing, China) and used for all experiments. Mice of similar age, regardless of sex, were randomly allocated to the experiments, as our previous study found no significant differences related to sex in light-induced retinal injury [ 30 ]. The mice were housed under a 12:12 h light/dark cycle with ad libitum access to standard mouse chow. Eye morphology was initially assessed as normal using a light microscope or fundus photography. When referring to 'the eye,' it denotes the eye of each individual mouse. The experimental design is illustrated in Fig. 1 A. Light exposure was conducted as previously described [ 30 ]. Mice were kept in constant darkness for 16 h prior to light exposure. Non-anesthetized mice were exposed to either typical laboratory lighting (50 lx) or white light (15,000 lx) for 2 h using, light-emitting diode (LED) lights positioned at the top of a standard mouse cage. LD was induced using LED desk lamps (Yeelight, model YLTD14YL, https://yeelight.tmall.com/shop/view_shop.htm ) with a color temperature of 5000 K, and light intensity was measured using a digital lux meter (TASI, model TA8120, Suzhou TASI Electronics Co., Ltd). Constant light exposure was performed between 14:00 and 16:00. Following light exposure, mice were allowed to recover for 2 h before receiving an intravitreal injection of TA or vehicle, as detailed in the Intravitreal drug administration section. Subsequent analyses were conducted one to three days after intravitreal injections; detailed information is provided in the figure legends. TA alleviates LD-induced impairment in retinal structure and function. A Schematic diagram illustrating the light exposure regimen and TA administration. BALB/c mice were dark-adapted for 16 h before being exposed to bright light (15,000 lx) for 2 h. Following light exposure, the mice were allowed to recover for 2 h before receiving an intravitreal (IV) injection of either vehicle or TA (15 μg/μL, 1 μL per eye). Analyses were performed one to three days after IV injection. B ERG results recorded two days after LD. Representative scotopic ERG responses to 0.1·cd s/m2 flashes from IV vehicle or IVTA-treated mice. C Luminance-response curves for the a- and b-waves from mice subjected to the indicated treatments. n = 6 mice per group. * p < 0.05, unpaired Student’s two-tailed t-test. D Upper panels: In vivo OCT images showing retinal structures. OCT was performed three days after LD. LD caused altered reflectance in the outer nuclear layer (ONL), which was mitigated in TA-treated mice. E HE staining showing retinal structures. Scale bar: 100 μm. n = 3 eyes per group. GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer. Lower panel: Quantification of ONL thickness. ONH: optic nerve head. # or * p < 0.05; #### or **** p < 0.0001 (#: control versus LD + vehicle; *: LD + vehicle versus LD + TA), two-way ANOVA, Tukey’s test. F TUNEL staining of the mouse retina after the indicated treatments. Mouse retinas were collected at the indicated time points following LD. Photoreceptor death was indicated by positive TUNEL staining, with DAPI counterstaining of nuclei. Scale bar: 100 μm. Right panel: Quantification of TUNEL-positive cells relative to the total number of DAPI-stained nuclei in the ONL. ns: not significant; *** p < 0.0005; n = 3 eyes per group. One-way ANOVA and Tukey’s test. For injection, mice were anesthetized with an intraperitoneal injection of 1% pentobarbital sodium (70 μL/10 g, prepared in normal saline solution). 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