ABSTRACT

Purpose

The gaseous signalling molecule, hydrogen sulfide (H₂S) has antioxidant, anti-inflammatory and anti-apoptotic properties. Since oxidative stress has been implicated in the pathogenesis of cataracts and lenticular hydrogen peroxide (H₂O₂) is elevated in some cataract patients, the present study investigated the ability of H₂S-releasing compounds to prevent H₂O₂-induced cataract formation in cultured bovine lenses.

Methods

Lenses were cultured in either Dulbecco's Modified Eagle Medium (DMEM; control); H₂O₂ (50 mM); ascorbic acid (AA; 3 mM) (positive control); and the H₂S-releasing compounds (diallyl trisulfide [DATS] or GYY4137) in the presence of H₂O₂ (50 mM). Lens opacity was determined using a plate reader to measure transmittance. Lens glutathione content (GSH), superoxide dismutase (SOD) activity and lactate dehydrogenase (LDH) cytotoxicity were assessed before and after treatment with the H₂S-releasing compounds.

Results

Both DATS (10⁻⁷M – 10⁻⁴M) and GYY4137 (10⁻⁷M – 10⁻⁴M) significantly (p < .001) attenuated H₂O₂ (50 mM)-induced loss in transmittance, with DATS (10⁻⁴M) and GYY4137 (10⁻⁷M) achieving a maximal reversal of opacity by 56.86 ± 0.01% (n = 6) and 8.39 ± 0.11% (n = 6) after 120 hours, respectively. These observations were corroborated by photographic evaluation, where DATS (10⁻⁵M – 10⁻⁴M) and GYY4137 (10⁻⁷M – 10⁻⁵M)-treated lenses had relatively clear grids after 120 hours, compared to H₂O₂ (50 mM)-treated lenses. The H₂O₂ (50 mM)-induced decline in total GSH content and total SOD activity were significantly (p < .001; n = 5) reversed by DATS (10⁻⁴M) and GYY4137 (10⁻⁷M). After 24 hours, DATS (10⁻⁴M) and GYY4137 (10⁻⁷M) significantly (p < .001; n = 4) reduced cytotoxicity of primary bovine lens epithelial cells by 33.88 ± 4.59% and 36.19 ± 10.53%, respectively.

Conclusion

Both H₂S-releasing compounds protected cultured bovine lenses against oxidative stress-induced cataract formation. The slow-releasing H₂S compound, GYY4137 was more potent than DATS in restoring lenticular total GSH content and total SOD activity along with reducing H₂O₂ (50 mM)-induced cytotoxicity

中文翻译：

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硫化氢释放化合物对培养牛晶状体中氧化应激诱导的白内障形成的保护作用

摘要

目的

气体信号分子硫化氢 (H ₂ S) 具有抗氧化、抗炎和抗凋亡特性。由于氧化应激与白内障的发病机制有关，一些白内障患者的晶状体过氧化氢 (H ₂ O ₂ ) 升高，本研究调查了释放H _{2 S 的化合物预防 H 2} O ₂诱导的白内障的能力在培养的牛晶状体中形成。

方法

镜片在 Dulbecco's Modified Eagle 培养基（DMEM；对照）中培养；H ₂ O ₂ (50 毫米); 抗坏血酸（AA；3 mM）（阳性对照）；和H ₂ O ₂ (50 mM)存在下的H _{2 S 释放化合物 (二烯丙基三硫化物 [DATS] 或 GYY4137)。}使用读板器测定透镜不透明度以测量透射率。在用释放H ₂ S 的化合物处理之前和之后评估晶状体谷胱甘肽含量 (GSH)、超氧化物歧化酶 (SOD) 活性和乳酸脱氢酶 (LDH) 细胞毒性。

结果

DATS (10 ^-7 M – 10 ^-4 M) 和 GYY4137 (10 ^-7 M – 10 ^-4 M) 均显着 ( p < .001) 减弱了 H ₂ O ₂ (50 mM) 引起的透射率损失，使用 DATS (10 ^-4 M) 和 GYY4137 (10 ^-7 M) 在 120 小时后分别实现了 56.86 ± 0.01% (n = 6) 和 8.39 ± 0.11% (n = 6) 的最大逆转。摄影评估证实了这些观察结果，其中 DATS (10 ^-5 M – 10 ^-4 M) 和 GYY4137 (10 ^-7 M – 10 ^-5 M) 处理的镜片在 120 小时后具有相对清晰的网格，与 H _2相比O ₂ (50 mM) 处理过的镜片。DATS (10 ^-4 M) 和 GYY4137 (10 ^-7 M) 可显着逆转H ₂ O ₂ (50 mM) 引起的总 GSH 含量和总 SOD 活性下降 ( p < .001；n = 5 )。24 小时后，DATS (10 ^-4 M) 和 GYY4137 (10 ^-7 M) 显着 ( p < .001；n = 4) 将原代牛晶状体上皮细胞的细胞毒性分别降低了 33.88 ± 4.59% 和 36.19 ± 10.53% .

结论

两种释放 H ₂ S 的化合物都能保护培养的牛晶状体免受氧化应激诱导的白内障形成。缓释 H ₂ S 化合物 GYY4137 在恢复透镜状总 GSH 含量和总 SOD 活性以及降低 H ₂ O ₂ (50 mM) 诱导的细胞毒性方面比 DATS 更有效