Abstract
Purpose
There is evidence that substrates for the endogenous production of hydrogen sulfide (H2S) such as L‐cysteine and N‐acetylcysteine (Zhang et al., Mol Vis, 14:862–870, 2008) and NSAIDs such as ibuprofen (Robert and Harding, Exp Eye Res, 54:509–518) can mitigate cataractogenesis in vitro and in vivo. However, the role of the hybrid compound consisting of the NSAID, indomethacin and a H2S‐donating moiety, ATB 343 on cataractogenesis has not been elucidated. In this study, we investigated the effect of the ATB 343 on cataractogenesis in cultured bovine lenses.
Methods
Freshly isolated bovine lenses were cultured in a DMEM buffer solution as follows: Group I: Control (DMEM); Group II–III: H2O2 (10 mM or 50 mM); Group IV–VI: ATB 343 (10−8M to 10−6M; Group VII: ascorbic acid (AA; 10 mM) in presence and absence of H2O2 (10 mM or 50 mM). Lenses were incubated in a CO2 chamber and assessed at 3, 6, 24, 48 and 72 hour‐time points. Qualitative assessments were conducted by visual inspection of lenses and photographic images captured against a black grid, while quantitative assessment was conducted by measurement of transmittance using a plate reader (Synergy H1 hybrid reader; Bio Tek Instruments, Inc) at every time point.
Results
DMEM‐cultured lenses exhibited a time‐dependent decrease in transmittance (420nm) and a corresponding loss of lens optical clarity up to 72 hours. Similar to the endogenous antioxidant, AA (10 mM), ATB 343 (10−7M) attenuated time‐dependent lens degradation up to 24 hours. H2O2 (10 mM and 50 mM) decreased light transmittance in a time‐dependent manner, achieving an inhibition of 41.98% (n=12; p<0.0001) and 46.37% (n=12; p<0.0001), respectively after 72 hours. Interestingly, ATB 343 (10−8 M to 10−6 M) partially reversed H2O2 (10 mM)‐induced degradation up to 72 hours. After 48 hours, the rank order of protection was: ATB (10−7M)>ATB (10−6M)>AA (10 mM)>ATB (10−8M). Similarly, ATB 343 (10−7 M and 10−6M) partially reversed H2O2 (50 mM)‐induced lens degradation up to 48 hours. After 24 hours, the rank order of protection was: ATB (10−7M)>ATB (10−8 M)>ATB (10−6M >AA (10 mM).
Conclusion
ATB 343 protected cultured lenses from time‐dependent degradation up to 24 hours and partially protected cultured bovine lenses from H2O2‐induced cataractogenesis.
Support or Funding Information
Creighton University‐SPAHP, Pharmaceutical Sciences Graduate Program
This is from the Experimental Biology 2019 Meeting. There is no full text article associated with this published in The FASEB Journal.