Bupivacaine Induces Reactive Oxygen Species Production via Activation of the AMP-Activated Protein Kinase-Dependent Pathway

Abstract

Aims: It was our aim to investigate whether AMP-activated protein kinase (AMPK) mediates the considerable increase in reactive oxygen species (ROS) and cell apoptosis induced by bupivacaine in the human neuroblastoma cell line SH-SY5Y. Methods: The recombinant plasmids pGPU6/GFP/Neo-shRNA AMPKα2 and pEGFP-N1-AMPKα2 were constructed and transfected into the SH-SY5Y cell line. The expression of AMPKα2 was determined by RT-PCR and Western blot after transfection. The SH-SY5Y cells transfected with recombinant plasmid were exposed to 1 mmol/l bupivacaine. Cell viability, intracellular ROS and apoptosis were determined. Results: The plasmid pEGFP-N1-AMPKα2 can upregulate the expression of AMPKα2, and the pGPU6/GFP/Neo-shRNA AMPKα2 can downregulate the expression of AMPKα2 in cells. Inhibition of AMPKα2 expression attenuated ROS production and cell apoptosis, and overexpression of AMPKα2 promoted ROS production and cell apoptosis after bupivacaine treatment. Conclusion: AMPK probably mediated ROS production and cell apoptosis induced by bupivacaine.

Autophagy activated by tuberin/mTOR/p70S6K suppression is a protective mechanism against local anaesthetics neurotoxicity

“…SH‐SY5Y cells were treated with bupivacaine, lidocaine, tetracaine, procaine and ropivacaine at different dosages according to the previous studies 7, 18, 19, 21, 22, 23, 24. Cell viability was evaluated by MTT assay 24 hrs after LAs challenge.…”

“…To address these questions, we treated human neuroblastoma SH‐SY5Y cells with both lipid LAs (procaine and tetracaine) and amide LAs (bupivacaine, lidocaine and ropivacaine) at previously described dosages 18, 19, 21, 22, 23, 24. The results showed that all the examined LAs up‐regulated autophagic process and inhibited tuberin/mTOR/p70S6K signalling in neuronal cells.…”

Therapeutic effects of intrathecal versus intravenous monosialoganglioside against bupivacaine-induced spinal neurotoxicity in rats

“…Excessive reactive oxygen species (ROS) causing neural injuries has also been proposed as another mechanism for bupivacaine-induce apoptosis. Bupivacaine induces intracellular accumulation of ROS which not only directly impairs neurons but also indirectly triggers apoptosis by activation of p38 AMP-activated protein kinase [24].…”

The Mechanisms Underlying Lipid Resuscitation Therapy

“…26 Whereas acute toxicity drives loss of insulinergic signaling, bathing cells overnight in bupivacaine induces cell death through apoptotic signaling at translational and mitochondrial targets downstream of Akt and mTOR. [27][28][29] In addition to loss of growth-related signaling, local anesthetics exert direct independent actions, which appreciably perturb the myocardial contractility apparatus. 30 It is unclear if these effects are the result of altered signaling at the membrane or an independent action.…”

“…Local anesthetics modify signaling via other kinases and second messengers including p38 MAPK, 120,121 AMPK, 29 ROS, 122 calcium sensitization, 123 protein kinase α, 124 and PKC. 124,125 Lipid also modifies signaling in many pathways while functioning as a metabolic fuel.…”