Enhanced Expression of WD Repeat-Containing Protein 35 via CaMKK/AMPK Activation in Bupivacaine-Treated Neuro2a Cells

Huang, Lei; Kondo, Fumio; Gosho, Masahiko; Feng, Guo-Gang; Harato, Misako; Xia, Zhongyuan; Ishikawa, Naohisa; Fujiwara, Yoshihiro; Okada, Shoshiro

doi:10.1371/journal.pone.0098185

Cited by 9 publications

(10 citation statements)

References 34 publications

(48 reference statements)

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“…AMPK, GSK-3β) occur during induction of toxicity and recovery from it. Our results agree with previous in vitro studies wherein lethal or cytotoxic concentrations of bupivacaine induce de-phosphorylation of Akt 7–9 and phosphorylation of AMPK 11,12 . Both of these kinases integrate signaling at mTORC1 to modulate sensitivity to endogenous insulin 30 during recovery from toxicity.…”

Section: Discussionsupporting

confidence: 93%

“…Specifically, 5’-adenosine-monophosphate kinase (AMPK) provides a countervailing force at tuberous sclerosis 2 (TSC2) that inhibits mTORC1 and downstream targets (Figure 2A) and has been implicated in cellular models of bupivacaine toxicity 11,12 . We probed for phosphorylation of AMPK at T172 and its downstream targets, Acetyl CoA Carboxylase (ACC) at S79 and TSC2 at S1387 (Figure 2B).…”

Section: Resultsmentioning

confidence: 99%

“…Further, the amide-linked local anesthetics, ropivacaine and lidocaine, disrupt assembly of phosphoinositide-3-kinase (Pi3k) thereby blocking phosphorylation of Akt 9 , an effect that is independent of classical sodium-channel blockade 10 . Beyond Akt, bupivacaine activates other controllers of glucose homeostasis, including 5’ adenosine monophosphate activated protein kinase (AMPK) 11,12 , an effect which may provide cyto-protection during toxicity 13 . Conversely, ILE and other fatty-acids increase phosphorylation of Akt and other canonical insulinergic targets when used as an adjuvant in recovery from ischemia-reperfusion injury 14–18 .…”

Section: Introductionmentioning

confidence: 99%

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Insulin Signaling in Bupivacaine-induced Cardiac Toxicity

et al. 2016

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Background The impact of local anesthetics on the regulation of glucose homeostasis by protein kinase B (Akt) and 5′-adenosine monophosphate–activated protein kinase (AMPK) is unclear but important because of the implications for both local anesthetic toxicity and its reversal by IV lipid emulsion (ILE). Methods Sprague–Dawley rats received 10 mg/kg bupivacaine over 20 s followed by nothing or 10 ml/kg ILE (or ILE without bupivacaine). At key time points, heart and kidney were excised. Glycogen content and phosphorylation levels of Akt, p70 s6 kinase, s6, insulin receptor substrate-1, glycogen synthase kinase-3β, AMPK, acetyl-CoA carboxylase, and tuberous sclerosis 2 were quantified. Three animals received Wortmannin to irreversibly inhibit phosphoinositide-3-kinase (Pi3k) signaling. Isolated heart studies were conducted with bupivacaine and LY294002—a reversible Pi3K inhibitor. Results Bupivacaine cardiotoxicity rapidly dephosphorylated Akt at S473 to 63 ± 5% of baseline and phosphorylated AMPK to 151 ± 19%. AMPK activation inhibited targets downstream of mammalian target of rapamycin complex 1 via tuberous sclerosis 2. Feedback dephosphorylation of IRS1 to 31 ± 8% of baseline sensitized Akt signaling in hearts resulting in hyperphosphorylation of Akt at T308 and glycogen synthase kinase-3β to 390 ± 64% and 293 ± 50% of baseline, respectively. Glycogen accumulated to 142 ± 7% of baseline. Irreversible inhibition of Pi3k upstream of Akt exacerbated bupivacaine cardiotoxicity, whereas pretreating with a reversible inhibitor delayed the onset of toxicity. ILE rapidly phosphorylated Akt at S473 and T308 to 150 ± 23% and 167 ± 10% of baseline, respectively, but did not interfere with AMPK or targets of mammalian target of rapamycin complex 1. Conclusion Glucose handling by Akt and AMPK is integral to recovery from bupivacaine cardiotoxicity and modulation of these pathways by ILE contributes to lipid resuscitation.

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Section: Discussionsupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Insulin Signaling in Bupivacaine-induced Cardiac Toxicity

et al. 2016

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“…25 ROS are known to stimulate a number of pathways that lead to cell death, including MAPK signal transduction pathways. Generally, p38 MAPK is preferentially stimulated by environmental stress and inflammatory cytokines 44,45 Huang et al showed that bupivacaine induces ROS and p38 MAPK, up-regulates the expression of the transcription factor c-Jun (AP-1) and significantly increases NF- κ B presence in the nucleus in neuronal cells. However, the stimulation or inhibition of these pathways by local anesthetics has been shown to be cell and system dependent.…”

Section: Discussionmentioning

confidence: 99%

Effect of Local Anesthetics on Human Mesenchymal Stromal Cell Secretion

et al. 2015

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Anti-fibrotic and tissue regenerative mesenchymal stromal cell (MSC) properties are largely mediated by secreted cytokines and growth factors. MSCs are implanted to augment joint cartilage replacement and to treat diabetic ulcers and burn injuries simultaneously with local anesthetics, which reduce pain. However, the effect of anesthetics on therapeutic human MSC secretory function has not been evaluated. In order to assess the effect of local anesthetics on the MSC secretome, a panel of four anesthetics with different potencies — lidocaine, procaine, ropivacaine and bupivacaine — was evaluated. Since injured tissues secrete inflammatory cytokines, the effects of anesthetics on MSCs stimulated with tumor necrosis factor (TNF)-α and interferon (IFN)-γ were also measured. Dose dependent and anesthesia specific effects on cell viability, post exposure proliferation and secretory function were quantified using alamar blue reduction and immunoassays, respectively. Computational pathway analysis was performed to identify upstream regulators and molecular pathways likely associated with the effects of these chemicals on the MSC secretome. Our results indicated while neither lidocaine nor procaine greatly reduced unstimulated cell viability, ropivacaine and bupivacaine induced dose dependent viability decreases. This pattern was exaggerated in the simulated inflammatory environment. The reversibility of these effects after withdrawal of the anesthetics was attenuated for TNF-α/IFN-γ-stimulated MSCs exposed to ropivacaine and bupivacaine. In addition, secretome analysis indicated that constitutive secretion changes were clearly affected by both anesthetic alone and anesthetic plus TNFα/IFNγ cell stimulation, but the secretory pattern was drug specific and did not necessarily coincide with viability changes. Pathway analysis identified different intracellular regulators for stimulated and unstimulated MSCs. Within these groups, ropivacaine and bupivacaine appeared to act on MSCs similarly via the same regulatory mechanisms. Given the variable effect of local anesthetics on MSC viability and function, these studies underscore the need to evaluate MSC in the presence of medications, such as anesthetics, that are likely to accompany cell implantation.

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“…It has been proposed that p38 mitogen‐activated protein kinase (p38 MAPK), protein kinase B (Akt), caspase pathways and apoptosis signaling play vital roles in local anesthetics‐induced neurotoxicity . For instance, bupivacaine‐induced neurotoxicity has been demonstrated to be closely associated with the activation of p38 MAPK and apoptosis signaling …”

Section: Introductionmentioning

confidence: 99%

Paeoniflorin attenuated bupivacaine‐induced neurotoxicity in SH‐SY5Y cells via suppression of the p38 MAPK pathway

Chen

Wang

et al. 2018

J of Cellular Biochemistry

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Bupivacain, a common local anesthetic, can cause neurotoxicity and permanent neurological disorders. Paeoniflorin has been widely reported as a potential neuroprotective agent in neural injury models. However, the roles and molecular basis of paeoniflorin in bupivacaine‐induced neurotoxicity are still undefined. In the current study, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay was performed to detect cell viability. Apoptotic rate was measured through double‐staining of Annexin V‐FITC and propidium iodide on a flow cytometer. Western blot assay was carried out to examine the protein levels of p38 mitogen‐activated protein kinase (p38 MAPK), phosphorylated‐p38 MAPK (p‐p38 MAPK), Bcl‐2, and Bax. caspase‐3 activity was determined using a caspase‐3 activity assay kit. We found that paeoniflorin dose‐dependently attenuated bupivacaine‐induced viability inhibition and apoptosis in SH‐SY5Y cells. Moreover, paeoniflorin inhibited bupivacaine‐induced activation of p38 MAPK pathway in SH‐SY5Y cells. Paeoniflorin alone showed no significant effect on cell viability, apoptosis and p38 MAPK signaling in SH‐SY5Y cells. Inhibition of p38 MAPK signaling by SB203580 or small interfering RNA targeting p38 (si‐p38) abated bupivacaine‐induced viability inhibition and apoptosis in SH‐SY5Y cells. In conclusion, paeoniflorin alleviated bupivacaine‐induced neurotoxicity in SH‐SY5Y cells via suppression of the p38 MAPK pathway, highlighting the potential values of paeoniflorin in relieving bupivacaine‐induced neurotoxicity.

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Enhanced Expression of WD Repeat-Containing Protein 35 via CaMKK/AMPK Activation in Bupivacaine-Treated Neuro2a Cells

Cited by 9 publications

References 34 publications

Insulin Signaling in Bupivacaine-induced Cardiac Toxicity

Insulin Signaling in Bupivacaine-induced Cardiac Toxicity

Effect of Local Anesthetics on Human Mesenchymal Stromal Cell Secretion

Paeoniflorin attenuated bupivacaine‐induced neurotoxicity in SH‐SY5Y cells via suppression of the p38 MAPK pathway

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