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Meclizine Preconditioning Protects the Kidney Against Ischemia-Reperfusion Injury.

Kishi S, Campanholle G, Gohil VM, Perocchi F, Brooks CR, Morizane R, Sabbisetti V, Ichimura T, Mootha VK, Bonventre JV - EBioMedicine (2015)

Bottom Line: Potentiation of glycolytic metabolism and attenuation of mitochondrial respiration may decrease cell injury and reduce reactive oxygen species generation from the mitochondria.Kidney injury was significantly decreased in meclizine treated mice compared with vehicle group (p < 0.001).Meclizine upregulated glycolysis in glucose-containing media and reduced cellular ATP levels in galactose-containing media.

View Article: PubMed Central - PubMed

Affiliation: Renal Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.

ABSTRACT
Global or local ischemia contributes to the pathogenesis of acute kidney injury (AKI). Currently there are no specific therapies to prevent AKI. Potentiation of glycolytic metabolism and attenuation of mitochondrial respiration may decrease cell injury and reduce reactive oxygen species generation from the mitochondria. Meclizine, an over-the-counter anti-nausea and -dizziness drug, was identified in a 'nutrient-sensitized' chemical screen. Pretreatment with 100 mg/kg of meclizine, 17 h prior to ischemia protected mice from IRI. Serum creatinine levels at 24 h after IRI were 0.13 ± 0.06 mg/dl (sham, n = 3), 1.59 ± 0.10 mg/dl (vehicle, n = 8) and 0.89 ± 0.11 mg/dl (meclizine, n = 8). Kidney injury was significantly decreased in meclizine treated mice compared with vehicle group (p < 0.001). Protection was also seen when meclizine was administered 24 h prior to ischemia. Meclizine reduced inflammation, mitochondrial oxygen consumption, oxidative stress, mitochondrial fragmentation, and tubular injury. Meclizine preconditioned kidney tubular epithelial cells, exposed to blockade of glycolytic and oxidative metabolism with 2-deoxyglucose and NaCN, had reduced LDH and cytochrome c release. Meclizine upregulated glycolysis in glucose-containing media and reduced cellular ATP levels in galactose-containing media. Meclizine inhibited the Kennedy pathway and caused rapid accumulation of phosphoethanolamine. Phosphoethanolamine recapitulated meclizine-induced protection both in vitro and in vivo.

No MeSH data available.


Related in: MedlinePlus

Cellular phosphoethanolamine is increased by meclizine and recapitulated meclizine-induced protection. (A) Phosphoethanolamine levels in HK-2 cells treated with or without 25 μM of meclizine for 17 h (n = 3). (B) Cellular ATP levels in HK-2 cells cultured in 10% DMEM containing 10 mM galactose with or without 10 μM of ethanolamine (EA) for 17 h (n = 4). (C) Lactate production in HK-2 cells after treatment with or without 10 μM of EA for 17 h (n = 7). (D) LDH release from HK-2 cells treated with either 10 or 100 μM of EA for 17 h followed by 2 hr of chemical anoxia (n = 7). (E) LDH release from LLC-PK1 cells treated with 10 μM of EA for 17 h followed by 2 hr of chemical anoxia (n = 4). (F) BUN and (G) serum creatinine concentrations at 24 and 48 h after IRI treated 2 h before, just after clamp removal and skin closure 24 h after ischemia with vehicle (n = 5) or EA (n = 4). (H) Representative images of H&E and PAS-stained kidney sections 48 h after IRI. Original magnification 100 ×, scale bar = 100 μm. (I) Summary of the mechanisms proposed for meclizine-induced protective effects against ischemic injury. Meclizine inhibits phosphate cytidylyltransferase 2 (PCYT2) and causes an increase in cytosolic phosphoethanolamine, a central precursor in the Kennedy pathway. High levels of intracellular phosphoethanolamine inhibit mitochondrial respiration.**p < 0.01 and *p < 0.05. Statistical significance was determined using t test (A, B, C, F, G) or one-way ANOVA followed by Tukey's post-hoc test (D, E). The columns and error bars are the mean ± SEM.
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f0040: Cellular phosphoethanolamine is increased by meclizine and recapitulated meclizine-induced protection. (A) Phosphoethanolamine levels in HK-2 cells treated with or without 25 μM of meclizine for 17 h (n = 3). (B) Cellular ATP levels in HK-2 cells cultured in 10% DMEM containing 10 mM galactose with or without 10 μM of ethanolamine (EA) for 17 h (n = 4). (C) Lactate production in HK-2 cells after treatment with or without 10 μM of EA for 17 h (n = 7). (D) LDH release from HK-2 cells treated with either 10 or 100 μM of EA for 17 h followed by 2 hr of chemical anoxia (n = 7). (E) LDH release from LLC-PK1 cells treated with 10 μM of EA for 17 h followed by 2 hr of chemical anoxia (n = 4). (F) BUN and (G) serum creatinine concentrations at 24 and 48 h after IRI treated 2 h before, just after clamp removal and skin closure 24 h after ischemia with vehicle (n = 5) or EA (n = 4). (H) Representative images of H&E and PAS-stained kidney sections 48 h after IRI. Original magnification 100 ×, scale bar = 100 μm. (I) Summary of the mechanisms proposed for meclizine-induced protective effects against ischemic injury. Meclizine inhibits phosphate cytidylyltransferase 2 (PCYT2) and causes an increase in cytosolic phosphoethanolamine, a central precursor in the Kennedy pathway. High levels of intracellular phosphoethanolamine inhibit mitochondrial respiration.**p < 0.01 and *p < 0.05. Statistical significance was determined using t test (A, B, C, F, G) or one-way ANOVA followed by Tukey's post-hoc test (D, E). The columns and error bars are the mean ± SEM.

Mentions: It has been shown that meclizine attenuated mitochondrial respiration by targeting cytosolic phosphoethanolamine metabolism (Gohil et al., 2013; Modica-Napolitano and Renshaw, 2004). Therefore, we asked whether the protective effect of meclizine pretreatment on kidney IRI is mediated through this pathway. As shown in Fig. 7A, meclizine pretreatment increased intracellular phosphoethanolamine in HK-2 cells (6.0 ± 0.62 vs 2.1 ± 0.10 μM, p < 0.05). Ethanolamine pretreatment significantly decreased ATP levels when cells were cultured in galactose-containing medium (3.0 ± 0.48 vs 5.2 ± 0.54 nmol/mg protein, p < 0.05) and was associated with increased lactate production (37.57 ± 12.71 vs 24.50 ± 3.18 nmol/h/mg, although not statistically significant p = 0.3352) (Fig. 7B, C). Ethanolamine pretreatment also inhibited % LDH release in response to 1.5 mM of NaCN and 10 mM of 2-DG-induced chemical hypoxia in HK-2 cells (10 μM: 23.9 ± 4.6 vs 49.0 ± 4.2%, p < 0.01 and 100 μM: 28.9 ± 5.2 vs 49.0 ± 4.2%, p < 0.05) and LLC-PK1 cells (10 μM; 39.1 ± 1.84 vs 65.0 ± 4.39%, p < 0.05) (Fig. 7D, E). When ethanolamine was given 2 h before ischemia, immediately after and 24 h after reperfusion, there was a significant decrease in BUN levels in mice exposed to IRI when compared to the vehicle group (24 h: 88.23 ± 12.23 vs 133.6 ± 7.15 mg/dl, p < 0.05; 48 h: 84.13 ± 19.62 vs 150.8 ± 12.26 mg/dl, p < 0.05). Serum creatinine levels at 24 h after IRI were also lower (1.25 ± 0.24 vs 1.94 ± 0.09 mg/dl, p < 0.05) (Fig. 7F, G). Compared with vehicle-injected mice, the renal morphology of ethanolamine-injected mice showed less kidney injury (Fig. 7H). These data revealed that phosphoethanolamine recapitulated meclizine-induced protection both in vitro and in vivo.


Meclizine Preconditioning Protects the Kidney Against Ischemia-Reperfusion Injury.

Kishi S, Campanholle G, Gohil VM, Perocchi F, Brooks CR, Morizane R, Sabbisetti V, Ichimura T, Mootha VK, Bonventre JV - EBioMedicine (2015)

Cellular phosphoethanolamine is increased by meclizine and recapitulated meclizine-induced protection. (A) Phosphoethanolamine levels in HK-2 cells treated with or without 25 μM of meclizine for 17 h (n = 3). (B) Cellular ATP levels in HK-2 cells cultured in 10% DMEM containing 10 mM galactose with or without 10 μM of ethanolamine (EA) for 17 h (n = 4). (C) Lactate production in HK-2 cells after treatment with or without 10 μM of EA for 17 h (n = 7). (D) LDH release from HK-2 cells treated with either 10 or 100 μM of EA for 17 h followed by 2 hr of chemical anoxia (n = 7). (E) LDH release from LLC-PK1 cells treated with 10 μM of EA for 17 h followed by 2 hr of chemical anoxia (n = 4). (F) BUN and (G) serum creatinine concentrations at 24 and 48 h after IRI treated 2 h before, just after clamp removal and skin closure 24 h after ischemia with vehicle (n = 5) or EA (n = 4). (H) Representative images of H&E and PAS-stained kidney sections 48 h after IRI. Original magnification 100 ×, scale bar = 100 μm. (I) Summary of the mechanisms proposed for meclizine-induced protective effects against ischemic injury. Meclizine inhibits phosphate cytidylyltransferase 2 (PCYT2) and causes an increase in cytosolic phosphoethanolamine, a central precursor in the Kennedy pathway. High levels of intracellular phosphoethanolamine inhibit mitochondrial respiration.**p < 0.01 and *p < 0.05. Statistical significance was determined using t test (A, B, C, F, G) or one-way ANOVA followed by Tukey's post-hoc test (D, E). The columns and error bars are the mean ± SEM.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4588407&req=5

f0040: Cellular phosphoethanolamine is increased by meclizine and recapitulated meclizine-induced protection. (A) Phosphoethanolamine levels in HK-2 cells treated with or without 25 μM of meclizine for 17 h (n = 3). (B) Cellular ATP levels in HK-2 cells cultured in 10% DMEM containing 10 mM galactose with or without 10 μM of ethanolamine (EA) for 17 h (n = 4). (C) Lactate production in HK-2 cells after treatment with or without 10 μM of EA for 17 h (n = 7). (D) LDH release from HK-2 cells treated with either 10 or 100 μM of EA for 17 h followed by 2 hr of chemical anoxia (n = 7). (E) LDH release from LLC-PK1 cells treated with 10 μM of EA for 17 h followed by 2 hr of chemical anoxia (n = 4). (F) BUN and (G) serum creatinine concentrations at 24 and 48 h after IRI treated 2 h before, just after clamp removal and skin closure 24 h after ischemia with vehicle (n = 5) or EA (n = 4). (H) Representative images of H&E and PAS-stained kidney sections 48 h after IRI. Original magnification 100 ×, scale bar = 100 μm. (I) Summary of the mechanisms proposed for meclizine-induced protective effects against ischemic injury. Meclizine inhibits phosphate cytidylyltransferase 2 (PCYT2) and causes an increase in cytosolic phosphoethanolamine, a central precursor in the Kennedy pathway. High levels of intracellular phosphoethanolamine inhibit mitochondrial respiration.**p < 0.01 and *p < 0.05. Statistical significance was determined using t test (A, B, C, F, G) or one-way ANOVA followed by Tukey's post-hoc test (D, E). The columns and error bars are the mean ± SEM.
Mentions: It has been shown that meclizine attenuated mitochondrial respiration by targeting cytosolic phosphoethanolamine metabolism (Gohil et al., 2013; Modica-Napolitano and Renshaw, 2004). Therefore, we asked whether the protective effect of meclizine pretreatment on kidney IRI is mediated through this pathway. As shown in Fig. 7A, meclizine pretreatment increased intracellular phosphoethanolamine in HK-2 cells (6.0 ± 0.62 vs 2.1 ± 0.10 μM, p < 0.05). Ethanolamine pretreatment significantly decreased ATP levels when cells were cultured in galactose-containing medium (3.0 ± 0.48 vs 5.2 ± 0.54 nmol/mg protein, p < 0.05) and was associated with increased lactate production (37.57 ± 12.71 vs 24.50 ± 3.18 nmol/h/mg, although not statistically significant p = 0.3352) (Fig. 7B, C). Ethanolamine pretreatment also inhibited % LDH release in response to 1.5 mM of NaCN and 10 mM of 2-DG-induced chemical hypoxia in HK-2 cells (10 μM: 23.9 ± 4.6 vs 49.0 ± 4.2%, p < 0.01 and 100 μM: 28.9 ± 5.2 vs 49.0 ± 4.2%, p < 0.05) and LLC-PK1 cells (10 μM; 39.1 ± 1.84 vs 65.0 ± 4.39%, p < 0.05) (Fig. 7D, E). When ethanolamine was given 2 h before ischemia, immediately after and 24 h after reperfusion, there was a significant decrease in BUN levels in mice exposed to IRI when compared to the vehicle group (24 h: 88.23 ± 12.23 vs 133.6 ± 7.15 mg/dl, p < 0.05; 48 h: 84.13 ± 19.62 vs 150.8 ± 12.26 mg/dl, p < 0.05). Serum creatinine levels at 24 h after IRI were also lower (1.25 ± 0.24 vs 1.94 ± 0.09 mg/dl, p < 0.05) (Fig. 7F, G). Compared with vehicle-injected mice, the renal morphology of ethanolamine-injected mice showed less kidney injury (Fig. 7H). These data revealed that phosphoethanolamine recapitulated meclizine-induced protection both in vitro and in vivo.

Bottom Line: Potentiation of glycolytic metabolism and attenuation of mitochondrial respiration may decrease cell injury and reduce reactive oxygen species generation from the mitochondria.Kidney injury was significantly decreased in meclizine treated mice compared with vehicle group (p < 0.001).Meclizine upregulated glycolysis in glucose-containing media and reduced cellular ATP levels in galactose-containing media.

View Article: PubMed Central - PubMed

Affiliation: Renal Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.

ABSTRACT
Global or local ischemia contributes to the pathogenesis of acute kidney injury (AKI). Currently there are no specific therapies to prevent AKI. Potentiation of glycolytic metabolism and attenuation of mitochondrial respiration may decrease cell injury and reduce reactive oxygen species generation from the mitochondria. Meclizine, an over-the-counter anti-nausea and -dizziness drug, was identified in a 'nutrient-sensitized' chemical screen. Pretreatment with 100 mg/kg of meclizine, 17 h prior to ischemia protected mice from IRI. Serum creatinine levels at 24 h after IRI were 0.13 ± 0.06 mg/dl (sham, n = 3), 1.59 ± 0.10 mg/dl (vehicle, n = 8) and 0.89 ± 0.11 mg/dl (meclizine, n = 8). Kidney injury was significantly decreased in meclizine treated mice compared with vehicle group (p < 0.001). Protection was also seen when meclizine was administered 24 h prior to ischemia. Meclizine reduced inflammation, mitochondrial oxygen consumption, oxidative stress, mitochondrial fragmentation, and tubular injury. Meclizine preconditioned kidney tubular epithelial cells, exposed to blockade of glycolytic and oxidative metabolism with 2-deoxyglucose and NaCN, had reduced LDH and cytochrome c release. Meclizine upregulated glycolysis in glucose-containing media and reduced cellular ATP levels in galactose-containing media. Meclizine inhibited the Kennedy pathway and caused rapid accumulation of phosphoethanolamine. Phosphoethanolamine recapitulated meclizine-induced protection both in vitro and in vivo.

No MeSH data available.


Related in: MedlinePlus