Chronic nicotine exposure augments renal oxidative stress and injury through transcriptional activation of p66shc

Arany, István; Clark, Jeb S.; Reed, Dustin K.; Juncos, Luis A.

doi:10.1093/ndt/gfs596

Cited by 48 publications

(74 citation statements)

References 42 publications

(89 reference statements)

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“…NRK52E cells were treated with either 200 μM NIC or 100 nM AZA or with their combination and LDH release was determined 24 h later. Figure 1A shows that both AZA and NIC increased LDH release similar to that found in a mouse proximal tubule cell line (8,18). Importantly, combination of NIC and AZA was additive.…”

Section: Resultssupporting

confidence: 70%

“…The result is enhanced mitochondrial ROS production and consequent mitochondrial depolarizationdependent injury (2). We revealed similar mechanism of injury by NIC treatment in renal proximal tubule cells (8). In this study, we determined that AZA-dependent toxicity is exacerbated by NIC, which is p66shc-and oxidative stressdependent ( Figure 1A).…”

Section: Discussionsupporting

confidence: 62%

“…p66sch is known as a pro-oxidant enzyme that links oxidative stress to renal injury (2,8,23,24). Interestingly, lately some studies -including ours (17)-revealed that under certain circumstances p66shc can serve as an antioxidant (25).…”

Section: Discussionmentioning

confidence: 86%

“…Hence, it is not surprising that knockdown of p66shc (shp66shc) attenuated NIC+AZA-associated increase in LDH release ( Figure 1A). Since p66shc exerts its cytotoxic effect via increasing oxidative stress (2,8), we determined whether NIC+AZA will do the same. Accordingly, cells were pretreated with the antioxidant N-acetyl-cysteine (NAC; 100 μM) and NIC+AZA-mediated LDH release was determined.…”

Section: Resultsmentioning

confidence: 99%

“…We also proved that adverse effect of NIC is due to augmented transcription of the p66shc gene ( Figure 1B). Further studies are needed to determine the mechanism of this activation, which could involve p53 and epigenetic changes in the p66shc promoter (2,8).…”

Section: Discussionmentioning

confidence: 99%

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Nicotine Exposure Augments Renal Toxicity of 5-azacytidine Through p66shc: Prevention by Resveratrol

Arany

Hall

Faisal

et al. 2017

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Abstract. Background Studies demonstrated that nephrotoxicity is a frequent side-effect of various anticancer agents that hampers their clinical use (1). Previously we reported that the anticancer agent 5-aza-cytidine (AZA) elicits oxidative stress-mediated toxicity in cultured renal proximal tubule cells via transcriptional upregulation of the prooxidant p66shc gene and consequent increase in mitochondrial reactive oxygen species (ROS) release (2). Studies revealed that smoking augments severity and progression of renal diseases in humans (3) and in various animal models (4-6). However, there are no data available on whether smoking could affect renal toxicity of anticancer agents.Smoking exerts its adverse effects via nicotine (NIC)-induced oxidative stress (7). We reported that NIC transcriptionally activates and Serine 36 phosphorylates p66shc that results in its mitochondrial translocation and therein cytochrome c binding and consequent excess ROS production (8) similar to AZA (2). Hence, it is plausible that NIC exposure exacerbates renal toxicity of AZA by further augmenting p66shc expression and resultant ROS production.Resveratrol (RES) is a powerful dietary antioxidant (9) that has been shown to protect the kidney from cigarette smoke-associated injury in rats (10). RES exerts antioxidant effect -at least partly -through activation of the Nrf2/HO-1 axis (11,12). Whether beneficial effect of RES on NIC+AZA-exposed renal cells are due to activation of Nrf2/HO-1 and/or down-regulation of p66shc is unknown.Accordingly, we hypothesized that (i) NIC exposure additively augments AZA-dependent induction of p66shc transcription, thus, exacerbates AZA-associated injury and (ii) RES attenuates NIC+AZA-mediated injury by suppressing p66shc transcription and/or activation of HO-1 in cultured renal proximal tubule cells. Materials and MethodsCell culture and treatment. The rat proximal tubule cell line (NRK52E) was maintained in 5% CO 2 at 37˚C in DMEM with 10% FBS as recommended. A set of cultures was treated with 200 μM nicotine (NIC; Sigma-Aldrich, St. Louis, MO, USA) or 100 nM 5-aza-cytidine (AZA; Sigma-Aldrich) as described earlier in mouse proximal tubule cells (2). Some cultures were co-treated with NIC+AZA. Resveratrol (RES; Selleckchem, Houston, TX, USA) was applied in 20 μM concentration overnight prior to NIC+AZA treatment.Determination of cell injury. Extent of cell injury was determined by LDH release using the fluorescent "Cytotox-One Homogeneous Membrane Integrity" kit (Promega, Madison, WI) as described earlier (13). Briefly, cells grown in 96-well-plates were transfected and treated as described in the appropriate section and 24 h later LDH content of the supernatants were determined and compared to total 4075

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Section: Resultssupporting

confidence: 70%

Section: Discussionsupporting

confidence: 62%

Section: Discussionmentioning

confidence: 86%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 3 more Smart Citations

Nicotine Exposure Augments Renal Toxicity of 5-azacytidine Through p66shc: Prevention by Resveratrol

Arany

Hall

Faisal

et al. 2017

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Measuring p66Shc Signaling Pathway Activation and Mitochondrial Translocation in Cultured Cells

et al. 2015

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The adaptor protein p66Shc links membrane receptors to intracellular signaling pathways, with downstream consequences on mitochondrial metabolism and reactive oxygen species production. Moreover, p66Shc has also been implicated in cancer development, progression, and metastasis. Increased phosphorylation of serine 36 residue of p66Shc very often correlates with oxidative stress-associated pathologies. The pro-oxidative role of p66Shc also appears to be involved in chemical toxicity, being an important component of stress responses triggered by xenobiotics. Here, we present a protocol that can be used: (a) for isolation of mitochondrial, cytosolic, and mitochondrial-associated membrane fractions from adherent cells lines; (b) to perform p66Shc detection with specific antibodies in order to monitor its translocation between different cellular compartments in response to the oxidative stress; and (c) to modulate the p66Shc pathway with the use of pharmacological approaches or gene-silencing methods.

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Coenzyme Q10 protects renal proximal tubule cells against nicotine-induced apoptosis through induction of p66shc-dependent antioxidant responses

et al. 2016

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Chronic nicotine exposure (via smoking, E-cigarettes) increases oxidative stress in the kidney that sensitizes it to additional injury in experimental models and in the renal patient. The pro-apoptotic p66 protein-via serine36 phosphorylation that facilitates its mitochondrial translocation and therein cytochrome c binding-generates oxidative stress that leads to injury of renal proximal tubule cells during chronic nicotine exposure. Coenzyme Q10-a clinically safe antioxidant-has been used against nicotine/smoke extract-associated oxidative stress in various non-renal cells. This study explored the anti-oxidant/anti-apoptotic effect of Coenzyme Q10 on nicotine-induced oxidative stress and its impact on p66shc in cultured rat renal proximal tubule cells (NRK52E). We studied the anti-oxidant effect of 10 µM Coenzyme Q10 using various mutants of the p66shc gene and also determined the induction of selected anti-oxidant entities (antioxidant response element, promoter of the manganese superoxide dismutase gene) in reporter luciferase assay during oxidative stress induced by 200 µM nicotine. Our studies revealed that Coenzyme Q10 strongly inhibits nicotine-mediated production of reactive oxygen species and consequent apoptosis that requires serine36 phosphorylation but not mitochondrial translocation/cytochrome c binding of p66. While both nicotine and Coenzyme Q10 stimulates the p66shc promoter, only nicotine exposure results in mitochondrial translocation of p66. In contrast, the Coenzyme Q10-stimulated and non-mitochondrial p66 activates the anti-oxidant manganese superoxide dismutase promoter via the antioxidant response elements and hence, rescues cells from nicotine-induced oxidative stress and consequent apoptosis.

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Chronic nicotine exposure augments renal oxidative stress and injury through transcriptional activation of p66shc

Abstract: 21. Hartner A, Porst M, Gauer S et al. Glomerular osteopontin expression and macrophage infiltration in glomerulosclerosis of DOCA-salt rats.

Cited by 48 publications

References 42 publications

Nicotine Exposure Augments Renal Toxicity of 5-azacytidine Through p66shc: Prevention by Resveratrol

Nicotine Exposure Augments Renal Toxicity of 5-azacytidine Through p66shc: Prevention by Resveratrol

Measuring p66Shc Signaling Pathway Activation and Mitochondrial Translocation in Cultured Cells

Coenzyme Q10 protects renal proximal tubule cells against nicotine-induced apoptosis through induction of p66shc-dependent antioxidant responses

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