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Lidocaine depolarizes the mitochondrial membrane potential by intracellular alkalization in rat dorsal root ganglion neurons

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Abstract

Purpose

The mitochondrial membrane potential (ΔΨm) is an important factor for apoptosis, and it is produced by the proton electrochemical gradient (ΔµH+). Therefore, the intracellular proton concentration (pHin) is an important factor for modifying the ΔΨm. However, the effects of lidocaine on pHin are unclear. To investigate mitochondrial responses to lidocaine, therefore, we simultaneously measured pHin with ΔΨm, flavin adenine dinucleotide (FAD), and reduced form of nicotinamide adenine dinucleotide (NADH) fluorescence, and calculated the FAD/NADH ratio (redox ratio), the superoxide production in mitochondria.

Methods

Morphological change and early apoptosis were observed by annexin-V FITC staining under fluorescent microscope. The ratiometric fluorescent probe JC-1 and HPTS were used for the simultaneous measurements of ΔΨm with pHin in rat dorsal root ganglion (DRG) neurons. FAD and NADH autofluorescence were simultaneously measured, and the FAD/NADH fluorescence ratio (redox ratio) was calculated. The superoxide was measured by mitosox-red fluorescent probe for mitochondrial superoxide. Lidocaine was evaluated at 1, 5, and 10 mM.

Results

Morphological change and early apoptosis were observed after 10 mM lidocaine administration. Lidocaine depolarized ΔΨm with increased pHin in a dose-dependent manner. In low-pH saline (pH 6), in the presence of both the weak acids (acetate and propionate), lidocaine failed to depolarize ΔΨm and increase pHin. On the other hand, lidocaine decreased the redox ratio in the cell and increased the levels of superoxide in a dose-dependent manner.

Conclusion

These results demonstrated that lidocaine depolarizes ΔΨm by intracellular alkalization. These results may indicate one of the mechanisms responsible for lidocaine-induced neurotoxicity.

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Acknowledgments

This work was supported in part by a Grant-in-Aid (No. 12770828) for Scientific Research (A) from The Ministry of Education, Science and Technology of Japan.

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Authors and Affiliations

  1. Department of Anesthesiology, Faculty of Medicine, University of Miyazaki, Kiyotake, Miyazaki, 889-1692, Japan

    Shin Onizuka, Tetsu Yonaha, Ryuji Tamura, Masatoshi Kasiwada, Toshiro Shirasaka & Isao Tsuneyoshi

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  1. Shin Onizuka
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  2. Tetsu Yonaha
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  3. Ryuji Tamura
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Correspondence to Shin Onizuka.

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Onizuka, S., Yonaha, T., Tamura, R. et al. Lidocaine depolarizes the mitochondrial membrane potential by intracellular alkalization in rat dorsal root ganglion neurons. J Anesth 25, 229–239 (2011). https://doi.org/10.1007/s00540-010-1079-y

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  • DOI: https://doi.org/10.1007/s00540-010-1079-y

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