David M Small, Nigel C Bennett, Sandrine Roy, Brian G Gabrielli, David W Johnson, Glenda C Gobe
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Adenosine triphosphate (ATP) and oxidative stress were measured by ATP assay, lipid peroxidation, total antioxidant capacity, mitochondrial function with confocal microscopy, MitoTracker Red CMXRos and live cell imaging with JC-1. In parallel, cell death and injury (i.e. apoptosis and Bax/Bcl-XL expression, lactate dehydrogenase), cell senescence (SA-β-galactosidase) and renal regenerative ability (cell proliferation), and their modulation with the anti-oxidant N-acetyl-cysteine (NAC) were investigated.</p><p><strong>Results: </strong>H2O2 and API, separately, increased oxidative stress and mitochondrial dysfunction, apoptosis and cell senescence. Although API caused cell senescence, it also induced oxidative stress at levels similar to H2O2 treatment alone, indicating that senescence and oxidative stress may be intrinsically linked. When H2O2 and API were delivered concurrently, their detrimental effects on renal cell loss were compounded. The antioxidant NAC attenuated apoptosis and senescence, and restored regenerative potential to the kidney.</p><p><strong>Conclusion: </strong>Oxidative stress and cell senescence both cause mitochondrial destabilization and cell loss and contribute to the development of the cellular characteristics of CKD.</p>","PeriodicalId":18993,"journal":{"name":"Nephron Experimental Nephrology","volume":"122 3-4","pages":"123-30"},"PeriodicalIF":0.0000,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000350726","citationCount":"55","resultStr":"{\"title\":\"Oxidative stress and cell senescence combine to cause maximal renal tubular epithelial cell dysfunction and loss in an in vitro model of kidney disease.\",\"authors\":\"David M Small, Nigel C Bennett, Sandrine Roy, Brian G Gabrielli, David W Johnson, Glenda C Gobe\",\"doi\":\"10.1159/000350726\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>The incidence and cost of chronic kidney disease (CKD) are increasing. 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In parallel, cell death and injury (i.e. apoptosis and Bax/Bcl-XL expression, lactate dehydrogenase), cell senescence (SA-β-galactosidase) and renal regenerative ability (cell proliferation), and their modulation with the anti-oxidant N-acetyl-cysteine (NAC) were investigated.</p><p><strong>Results: </strong>H2O2 and API, separately, increased oxidative stress and mitochondrial dysfunction, apoptosis and cell senescence. Although API caused cell senescence, it also induced oxidative stress at levels similar to H2O2 treatment alone, indicating that senescence and oxidative stress may be intrinsically linked. When H2O2 and API were delivered concurrently, their detrimental effects on renal cell loss were compounded. 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引用次数: 55
摘要
背景:慢性肾脏疾病(CKD)的发病率和费用正在增加。肾小管上皮细胞功能障碍和损耗,包括细胞凋亡增加和细胞衰老,是CKD发病机制的核心。目的是使用体外模型来研究氧化应激、线粒体功能障碍和细胞衰老在促进肾肿块损失中的单独和累积作用。方法:用中度过氧化氢(H2O2)治疗人肾小管上皮细胞(HK2)的氧化应激,用或不加细胞周期抑制剂(apigenin, API)治疗细胞衰老。通过ATP测定、脂质过氧化、总抗氧化能力、共聚焦显微镜、MitoTracker Red CMXRos和JC-1活细胞成像检测三磷酸腺苷(ATP)和氧化应激。同时,研究了细胞死亡和损伤(即凋亡和Bax/Bcl-XL表达、乳酸脱氢酶)、细胞衰老(SA-β-半乳糖苷酶)和肾再生能力(细胞增殖),以及抗氧化剂n -乙酰半胱氨酸(NAC)对它们的调节作用。结果:H2O2和API分别增加氧化应激和线粒体功能障碍、细胞凋亡和细胞衰老。API虽然引起细胞衰老,但其诱导氧化应激的水平与单独处理H2O2相似,表明衰老与氧化应激可能存在内在联系。当H2O2和API同时递送时,它们对肾细胞损失的有害影响更加严重。抗氧化剂NAC可减轻细胞凋亡和衰老,恢复肾脏再生潜能。结论:氧化应激和细胞衰老均导致线粒体失稳和细胞丢失,参与CKD细胞特征的形成。
Oxidative stress and cell senescence combine to cause maximal renal tubular epithelial cell dysfunction and loss in an in vitro model of kidney disease.
Background: The incidence and cost of chronic kidney disease (CKD) are increasing. Renal tubular epithelial cell dysfunction and attrition, involving increased apoptosis and cell senescence, are central to the pathogenesis of CKD. The aim here was to use an in vitro model to investigate the separate and cumulative effects of oxidative stress, mitochondrial dysfunction and cell senescence in promoting loss of renal mass.
Methods: Human kidney tubular epithelial cells (HK2) were treated with moderate hydrogen peroxide (H2O2) for oxidative stress, with or without cell cycle inhibition (apigenin, API) for cell senescence. Adenosine triphosphate (ATP) and oxidative stress were measured by ATP assay, lipid peroxidation, total antioxidant capacity, mitochondrial function with confocal microscopy, MitoTracker Red CMXRos and live cell imaging with JC-1. In parallel, cell death and injury (i.e. apoptosis and Bax/Bcl-XL expression, lactate dehydrogenase), cell senescence (SA-β-galactosidase) and renal regenerative ability (cell proliferation), and their modulation with the anti-oxidant N-acetyl-cysteine (NAC) were investigated.
Results: H2O2 and API, separately, increased oxidative stress and mitochondrial dysfunction, apoptosis and cell senescence. Although API caused cell senescence, it also induced oxidative stress at levels similar to H2O2 treatment alone, indicating that senescence and oxidative stress may be intrinsically linked. When H2O2 and API were delivered concurrently, their detrimental effects on renal cell loss were compounded. The antioxidant NAC attenuated apoptosis and senescence, and restored regenerative potential to the kidney.
Conclusion: Oxidative stress and cell senescence both cause mitochondrial destabilization and cell loss and contribute to the development of the cellular characteristics of CKD.