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. 2024 Jul 1;65(8):1.
doi: 10.1167/iovs.65.8.1.

Protective Effect of Nicotinamide Riboside on Glucocorticoid-Induced Glaucoma: Mitigating Mitochondrial Damage and Extracellular Matrix Deposition

Nan Zhang  1 , Pengyu Zhang  1 , Xizhi Deng  1 , Min Zhu  1 , Yixin Hu  1 , Dongxiao Ji  1   2 , Lufan Li  1 , Yang Liu  1 , Wen Zeng  1 , Min Ke  1
Affiliations

Protective Effect of Nicotinamide Riboside on Glucocorticoid-Induced Glaucoma: Mitigating Mitochondrial Damage and Extracellular Matrix Deposition

Nan Zhang et al. Invest Ophthalmol Vis Sci. .

Abstract

Purpose: Glucocorticoid-induced glaucoma (GIG) is a prevalent complication associated with glucocorticoids (GCs), resulting in irreversible blindness. GIG is characterized by the abnormal deposition of extracellular matrix (ECM) in the trabecular meshwork (TM), elevation of intraocular pressure (IOP), and loss of retinal ganglion cells (RGCs). The objective of this study is to investigate the effects of nicotinamide riboside (NR) on TM in GIG.

Methods: Primary human TM cells (pHTMs) and C57BL/6J mice responsive to GCs were utilized to establish in vitro and in vivo GIG models, respectively. The study assessed the expression of ECM-related proteins in TM and the functions of pHTMs to reflect the effects of NR. Mitochondrial morphology and function were also examined in the GIG cell model. GIG progression was monitored through IOP, RGCs, and mitochondrial morphology. Intracellular nicotinamide adenine dinucleotide (NAD+) levels of pHTMs were enzymatically assayed.

Results: NR significantly prevented the expression of ECM-related proteins and alleviated dysfunction in pHTMs after dexamethasone treatment. Importantly, NR protected damaged ATP synthesis, preventing overexpression of mitochondrial reactive oxygen species (ROS), and also protect against decreased mitochondrial membrane potential induced by GCs in vitro. In the GIG mouse model, NR partially prevented the elevation of IOP and the loss of RGCs. Furthermore, NR effectively suppressed the excessive expression of ECM-associated proteins and mitigated mitochondrial damage in vivo.

Conclusions: Based on the results, NR effectively enhances intracellular levels of NAD+, thereby mitigating abnormal ECM deposition and TM dysfunction in GIG by attenuating mitochondrial damage induced by GCs. Thus, NR has promising potential as a therapeutic candidate for GIG treatment.

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Conflict of interest statement

Disclosure: N. Zhang, None; P. Zhang, None; X. Deng, None; M. Zhu, None; Y. Hu, None; D. Ji, None; L. Li, None; Y. Liu, None; W. Zeng, None; M. Ke, None

Figures

Figure 1.
Figure 1.
Nicotinamide riboside (NR) prevented the overexpression of the extracellular matrix (ECM) induced by DEX. (A) Protein levels of ECM–related markers (fibronectin [FN], COL4, and Myocilin) were assessed through Western blotting, and quantifications were performed. Data presented are from six individual Western blot experiments, which were derived from three separate donor cultures. One-way ANOVA with Tukey's multiple tests was conducted for each ECM marker, * P < 0.05, ** P < 0.01, *** P < 0.001, no statistical difference for unmarked comparisons. (B) Representative images of pHTMs stained against FN and COL4A from various treated groups, along with quantification of fluorescence intensity (n ≥ 9 fields per group). The results are expressed as mean ± standard error of the mean. One-way ANOVA with Tukey's multiple tests was conducted, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Scale bar = 200 µm (B).
Figure 2.
Figure 2.
Nicotinamide riboside (NR) attenuated the dysfunction of primary human trabecular meshwork cells (pHTMs) induced by DEX in vitro. (A) Phagocytosis function of pHTMs was measured using flow cytometry. (B) Cell proliferation of pHTMs was evaluated by EdU staining. Representative images of pHTMs stained with EdU from different treated groups and quantifications of proliferative rates. (C) The Transwell test demonstrated that NR prevented DEX-induced increased migration of pHTMs. The results are represented as mean ± standard error of the mean. One-way ANOVA with Tukey's multiple tests, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. The data analyzed in Figure 3A were from 3 separate donor cultures (n ≥ 9 fields per groups in panels B and C). Data presented in Figures 3B and 3C represent at least three biological replicates from three separate donor cultures. Scale bar = 200 µm.
Figure 3.
Figure 3.
Nicotinamide riboside (NR) prevented mitochondrial damage in primary human trabecular meshwork cells (pHTMs) induced by DEX in vitro. (A) Mitochondrial morphology was assessed using MitoTracker dye (n ≥ 27 cells analyzed per groups). (B, C) Mitochondrial membrane potential (Ψm) was measured by tetramethylrhodamine ethyl ester perchlorate (B) and JC-1 staining (C). (D) Mitochondrial ROS (mtROS) was detected by MitoSOX Red Dye. (E) NR prevented damaged ATP production in the GIG cell model. (F) NR elevated intracellular NAD+ levels in the GIG cell model. The results are presented as mean ± standard error of the mean. One-way ANOVA with Tukey's multiple tests, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (n ≥ 12 fields per groups in panels B, C, and D). Data presented in B, C, and D represent at least three biological replicates from three separate donor cultures. Data presented in panels E and F are two replicated samples from three independent donor cultures, totaling six samples per group. Scale bar = 50 µm (A), 200 µm (B, C, D).
Figure 4.
Figure 4.
Nicotinamide riboside (NR) prevented the progression of GIG in vivo. (A) Schematic course of GIG mouse model building and NR treatment. (B) NR partially prevented increased intraocular pressure in the GIG mouse model (red asterisk = NC versus DEX-Ace, and the orange asterisk = DEX-Ace versus DEX-Ace+NR; n ≥ 12 eyes per groups). One-way ANOVA with Tukey's multiple tests was conducted for each time point, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. (C) NR protected against RGC loss in the GIG cell model. Representative images of retina flat-mounts stained against RBPMS from different treated groups. Images were randomly chosen from the region with a distance to the optic nerve of 1.0 mm (n = 6–8 retinas per group). One-way ANOVA with Tukey's multiple tests, ** P < 0.01, **** P < 0.0001. (D) NR depressed the overexpression of ECM-related proteins (FN and COL4A), as tested by Western blotting (n = 4 mice per group). One-way ANOVA with Tukey's multiple tests was conducted for each ECM marker, * P < 0.05, ** P < 0.01, *** P < 0.001, no statistical difference for unmarked comparisons. (E) Mitochondrias in the TM region were imaged by electron microscopy. Black arrows indicate damaged mitochondria in the DEX-Ace treated group. The results are expressed as mean ± standard error of the mean. Scale bar = 100 µm (C), 10 µm (upper E), 1 µm (middle E), and 500 nm (lower E).
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References

    1. Vandewalle J, Luypaert A, De Bosscher K, Libert C. Therapeutic mechanisms of glucocorticoids. Trends Endocrinol Metab. 2018; 29(1): 42–54. - PubMed
    1. Kato M, Hosoe N, Gotoda T, et al. .. Treatment with vonoprazan for 3 weeks is not inferior to 8 weeks for the management of gastric ESD: a multicenter noninferiority randomized study. J Gastroenterol. 2023; 58(4): 358–366. - PubMed
    1. Roy Chowdhury U, Hann CR, Stamer WD, Fautsch MP. Aqueous humor outflow: dynamics and disease. Invest Ophthalmol Vis Sci. 2015; 56(5): 2993–3003. - PMC - PubMed
    1. Fini ME, Schwartz SG, Gao X, et al. .. Steroid-induced ocular hypertension/glaucoma: focus on pharmacogenomics and implications for precision medicine. Prog Retin Eye Res. 2017; 56: 58–83. - PMC - PubMed
    1. Wang K, Read AT, Sulchek T, Ethier CR. Trabecular meshwork stiffness in glaucoma. Exp Eye Res. 2017; 158: 3–12. - PMC - PubMed

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