AP39, A Mitochondrially Targeted Hydrogen Sulfide Donor, Exerts Protective Effects in Renal Epithelial Cells Subjected to Oxidative Stress in Vitro and in Acute Renal Injury in Vivo.
AP39, A Mitochondrially Targeted Hydrogen Sulfide Donor, Exerts Protective Effects in Renal Epithelial Cells Subjected to Oxidative Stress in Vitro and in Acute Renal Injury in Vivo.
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DOI：
10.1097/shk.0000000000000478
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发表时间：
2016-01
期刊：
影响因子：
--
通讯作者：
Szabo C
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文献类型：
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作者：
Ahmad A;Olah G;Szczesny B;Wood ME;Whiteman M;Szabo C
本研究评估了线粒体靶向供体硫化氢（H2S） AP39[（10-氧-10-（4-（3-硫氧- 3h -1,2-二硫醇-5基）苯氧基）十基）三苯基溴化磷]在NRK- 49f大鼠肾上皮细胞（NRK细胞）缺氧/氧化应激损伤体外模型和大鼠肾缺血再灌注损伤模型中的作用。添加葡萄糖氧化酶可诱导肾脏氧化应激，葡萄糖氧化酶在培养基中以恒定速率生成过氧化氢。葡萄糖氧化酶（GOx）诱导的氧化应激导致线粒体功能障碍，降低细胞内ATP含量，并且在较高浓度下，细胞内氧化剂形成增加（通过荧光探针2,7 -二氯荧光素，DCF估计）并促进NRK细胞的坏死（通过测量乳酸脱氢酶释放到培养基中估计）。AP39预处理（30 ~ 300 nM）对GOx的上述作用均具有浓度依赖性的保护作用。AP39的大部分作用呈钟形浓度-响应曲线；在GOx浓度最高时，AP39不再具有细胞保护作用。肾缺血/再灌注大鼠的血尿素氮和肌酐水平明显升高（比对照基线高出4倍以上），表明肾脏严重受损。这与中性粒细胞向肾脏的浸润增加（通过肾匀浆中的髓过氧化物酶测定来评估）、氧化应激增加（通过肾匀浆中的丙二醛测定来评估）和血浆IL-12水平增加有关。AP39预处理（0.1、0.2和0.3 mg/kg）对这些病理生理改变具有剂量依赖性保护作用；以H2S供体剂量0.3 mg/kg时保护作用最显著；然而，AP39未能实现任何损伤指标的完全正常化。AP39的部分保护作用与肾脏组织学评分的部分改善和TUNEL染色（DNA损伤和凋亡的指标）的降低相关。综上所述，线粒体靶向H2S供体AP39对体外肾上皮细胞损伤和体内肾缺血再灌注损伤具有剂量依赖性的保护作用。我们假设AP39的有益作用与细胞氧化应激的减少以及随后炎症和氧化过程的各种正反馈周期的衰减有关。
This study evaluated the effects of AP39 [(10-oxo-10-(4-(3-thioxo-3H-1,2-dithiol-5yl) phenoxy)decyl) triphenyl phosphonium bromide], a mitochondrially targeted donor of hydrogen sulfide (H2S) in an in vitro model of hypoxia/oxidative stress injury in NRK-49F rat kidney epithelial cells (NRK cells) and in a rat model of renal ischemia-reperfusion injury. Renal oxidative stress was induced by the addition of glucose oxidase, which generates hydrogen peroxide in the culture medium at a constant rate. Glucose oxidase (GOx)-induced oxidative stress led to mitochondrial dysfunction, decreased intracellular ATP content, and, at higher concentrations, increased intracellular oxidant formation (estimated by the fluorescent probe 2, 7-dichlorofluorescein, DCF) and promoted necrosis (estimated by the measurement of lactate dehydrogenase release into the medium) of the NRK cells in vitro. Pretreatment with AP39 (30-300 nM) exerted a concentration-dependent protective effect against all of the above effects of GOx. Most of the effects of AP39 followed a bell-shaped concentration-response curve; at the highest concentration of GOx tested, AP39 was no longer able to afford cytoprotective effects. Rats subjected to renal ischemia/reperfusion responded with a marked increase (over 4-fold over sham control baseline) blood urea nitrogen and creatinine levels in blood, indicative of significant renal damage. This was associated with increased neutrophil infiltration into the kidneys (assessed by the myeloperoxidase assay in kidney homogenates), increased oxidative stress (assessed by the malondialdehyde assay in kidney homogenates) and an increase in plasma levels of IL-12. Pretreatment with AP39 (0.1, 0.2 and 0.3 mg/kg) provided a dose-dependent protection against these pathophysiological alterations; the most pronounced protective effect was observed at the 0.3 mg/kg dose of the H2S donor; nevertheless AP39 failed to achieve a complete normalization of any of the injury markers measured. The partial protective effects of AP39 correlated with a partial improvement of kidney histological scores and reduced TUNEL staining (an indicator of DNA damage and apoptosis). In summary, the mitochondria-targeted H2S donor AP39 exerted dose-dependent protective effects against renal epithelial cell injury in vitro and renal ischemia-reperfusion injury in vivo. We hypothesize that the beneficial actions of AP39 are related to the reduction of cellular oxidative stress, and subsequent attenuation of various positive feed-forward cycles of inflammatory and oxidative processes.