Regulation of Cataractogenesis by Hydrogen Sulfide-NSAID Hybrid Compounds in Cultured Bovine Lenses

Abstract

Purpose : Hydrogen sulfide (H₂S) releasing compounds and NSAIDs can mitigate cataractogenesis in vitro and in vivo (Zhang et al., Mol Vis, 14:862, 2008; Harding JJ et al., Acta Ophthalmol, 67:518, 1989). However, the role of NSAID-H₂S-releasing hybrid compounds on cataractogenesis has not been elucidated. In the present study, we compared the pharmacological effects of ATB343 and ATB337 on cataractogenesis in cultured bovine lenses and on lenticular antioxidants concentrations.

Methods : Freshly isolated bovine lenses were cultured in a DMEM buffer solution as follows: Group I: Control (DMEM); Group II: H₂O₂ (10 mM); Groups III-V: ATB343 (10^-7M); ATB337 (10^-7M); ascorbic acid (AA; 10^-3M) in presence and absence of H₂O₂. Lenses were incubated and were qualitatively and quantitatively assessed at 3, 6, 24, 48 and 72 h-time points by photographic captures and measurement of transmittance using a plate reader (Synergy H1 hybrid). Lenticular glutathione (GSH) and superoxide dismutase (SOD) were measured using Cayman Assay Kits

Results : DMEM-cultured lenses elicited a time-dependent decrease in transmittance (420nm) and a corresponding loss of lens optical clarity up to 72 h. Unlike ATB343 (10^-7M) and AA (10 mM) which attenuated time-dependent lens degradation up to 24 h, ATB337 (10^-7M) decreased time-dependent transmittance, achieving an inhibition of 34.7 ± 1.12% (n=12; p<0.01) after 72 h. H₂O₂ (10 mM) reduced transmittance in a time-dependent manner, attaining an inhibition of 42.0 ± 1.0% (n=12; p<0.01) after 72 h. After 48 h, ATB343 (10^-7M) and ATB337 (10^-7M) enhanced time-dependent GSH depletion by 3.6%± 0.05% (n=3; p<0.05) and 2.7± 0.5% (n=3; p<0.05) and H₂O₂-induced GSH depletion by 12.3± 0.1% (n=3; p<0.01) and 8.7 ±0.4% (n=3; p<0.05), respectively. However, ATB343 enhanced SOD depletion by 35.3 ± 2.7% (n=3; p<0.001) but attenuated H₂O₂-induced SOD depletion by 56.6±3.1% (n=3; p<0.001) (t=48 h). Similarly, ATB337 enhanced time-dependent loss in SOD by 48.9 ± 4.4% (n=3; p<0.001) but reversed H₂O₂-induced SOD loss by 31.8 ± 4.9% (n=3; p<0.001) (t=48 h).

Conclusions : The NSAID-H₂S hybrid compounds, ATB343 and ATB337 can partially protect cultured bovine lenses from cataract formation, presumably via an effect on lenticular SOD concentrations.

This is a 2020 ARVO Annual Meeting abstract.