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模拟皮质性白内障形成22:牛磺酸因其抗氧化活性在体外减轻模型糖尿病大鼠白内障损伤?

Modelling cortical cataractogenesis 22: is in vitro reduction of damage in model diabetic rat cataract by taurine due to its antioxidant activity?

作者信息

Kilic F, Bhardwaj R, Caulfeild J, Trevithick J R

机构信息

Department of Biochemistry, Faculty of Medicine and Dentistry, University of Western Ontario, London, Ontario, N6A 5C1, Canada.

出版信息

Exp Eye Res. 1999 Sep;69(3):291-300. doi: 10.1006/exer.1999.0697.

Abstract

The protective effect of taurine in model in vitro diabetic cataract and the mechanism of this effect were investigated in isolated rat lenses. Isolated rat lenses were incubated in medium 199 in elevated glucose (55.6 m m) with taurine (5 m m). Taurine concentrations in the lenses were determined by amino acid analysis. Accumulative leakage of the intracellular enzyme lactate dehydrogenase (LDH) was used to estimate damage to the lens, as previously reported. In the clear lenses, prior to vacuole formation, after 1 or 2 days of incubation, the taurine and amino acids in lenses decreased progressively in concentration. In lenses incubated with 5 m m taurine, the level of taurine was increased towards that of control lenses. In taurine-treated lenses LDH leakage was significantly decreased, and lens clarity was maintained, similarly to that found previously for vitamin C and lipoic acid. To test whether taurine has similar antioxidant activity, we tested its ability to decrease luminol luminescence generated by (1) superoxide from hypoxanthine/xanthine oxidase and (2) peroxide from diluted glucose/glucose oxidase. For either superoxide or peroxide, the luminescence was decreased to zero, as a function of increasing taurine concentration, at 30 m m, approximately the physiological concentration of taurine in the lens. Spin trapping confirmed that taurine scavenged superoxide. This is consistent with a role for taurine as an important antioxidant protecting the lens against oxidative insults. Amino acids also had antioxidant activity in this assay, and as a group, when all activities were summed, their loss also contributed significantly to the antioxidant loss. Taken in conjunction with Wolff and Crabbe's observation of increased free radical generation by glucose auto-oxidation in diabetes, this suggests a push-pull mechanism for increased oxidative stress in diabetic cataract, involving both increased free radicals and decreased radical scavenging antioxidants.

摘要

在离体大鼠晶状体中研究了牛磺酸对体外糖尿病性白内障模型的保护作用及其作用机制。将离体大鼠晶状体置于含55.6 mM高糖及5 mM牛磺酸的199培养基中孵育。通过氨基酸分析测定晶状体中的牛磺酸浓度。如先前报道,采用细胞内酶乳酸脱氢酶(LDH)的累积泄漏来评估晶状体损伤情况。在清亮晶状体中,空泡形成前,孵育1或2天后,晶状体中的牛磺酸和氨基酸浓度逐渐降低。在含5 mM牛磺酸孵育的晶状体中,牛磺酸水平向对照晶状体的水平升高。在牛磺酸处理的晶状体中,LDH泄漏显著减少,晶状体透明度得以维持,类似于先前维生素C和硫辛酸的情况。为测试牛磺酸是否具有类似的抗氧化活性,我们测试了其降低由(1)次黄嘌呤/黄嘌呤氧化酶产生的超氧化物和(2)稀释葡萄糖/葡萄糖氧化酶产生的过氧化物所引发的鲁米诺发光的能力。对于超氧化物或过氧化物,随着牛磺酸浓度增加,在约30 mM(晶状体中牛磺酸的生理浓度)时,发光降至零。自旋捕获证实牛磺酸清除了超氧化物。这与牛磺酸作为重要抗氧化剂保护晶状体免受氧化损伤的作用一致。在该测定中氨基酸也具有抗氧化活性,总体而言,当所有活性相加时,它们的损失也显著导致了抗氧化剂损失。结合Wolff和Crabbe关于糖尿病中葡萄糖自氧化导致自由基生成增加的观察结果,这提示糖尿病性白内障中氧化应激增加的一种推拉机制,涉及自由基增加和自由基清除抗氧化剂减少。

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