Effect of local anestbetic on neuronal cytoplasmic calcium and plasma membrane lysis (necrosis) in a cell culture model

被引:80
作者
Johnson, ME
Saenz, JA
DaSilva, AD
Uhl, CB
Gores, GJ
机构
[1] Mayo Clin & Mayo Fdn, Dept Anesthesiol, Rochester, MN 55905 USA
[2] Sch Med, Rochester, MN USA
关键词
D O I
10.1097/00000542-200212000-00019
中图分类号
R614 [麻醉学];
学科分类号
100217 ;
摘要
Background: To investigate the mechanism by which rare cases of spinal local anesthetic (LA) neurotoxicity occur, we have tested the hypotheses that LAs elevate cytoplasmic calcium (Ca-cyt(2+)), that this is associated with a neurotoxic effect, and that lidocaine and bupivacaine differ in their neurotoxicity. Methods: Neurons of the ND7 cell culture line, derived from dorsal root ganglion, were loaded with fura-2 and analyzed by digitized video fluorescence microscopy during 60 min LA exposure, allowing determination of Ca-cyt(2+) and time of necrotic cell death (plasma membrane lysis) at the single neuron level. Results: Lidocaine 0.1% and bupivacaine 0.025% caused minimal changes in Ca-cyt(2+). Lidocaine 0.5-5% and bupivacaine 0.125-0.625% caused an early, small (less than threefold), concentration-dependent increase in Ca-cyt(2+) that was transient and returned to near baseline within 10 min. Lidocaine 2.5% and 5% then caused a sustained, greater than ten-fold increase in Ca-cyt(2+) and death in some neurons during the 60 min exposure period. Pretreatment with thapsigargin eliminated the initial transient increase in Ca-cyt(2+) consistent with endoplasmic reticulum (ER) as its source, and increased neuronal death with 5% lidocaine, suggesting that lidocaine neurotoxicity can be increased by failure of ER to take up elevated Ca-cyt(2+). The later sustained increase in Ca-cyt(2+) seen with 2.5 and 5% lidocaine was prevented in Ca2+-free medium, and restored when Ca 21 was added back to the buffer in the presence of lidocaine, suggesting that higher concentrations of lidocaine increase influx of Ca2+ through the plasma membrane. Conclusions: In this model, lidocaine greater than 2.5% elevates Ca-cyt(2+) to toxic levels. Bupivacaine and lower concentrations of lidocaine transiently alter Ca-cyt(2+), homeostasis for several minutes, but without an immediate neurotoxic effect within 60 min.
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页码:1466 / 1476
页数:11
相关论文
共 46 条
[1]   Time domains of neuronal Ca2+ signaling and associative memory:: steps through a calexcitin, ryanodine receptor, K+ channel cascade [J].
Alkon, DL ;
Nelson, TJ ;
Zhao, WQ ;
Cavallaro, S .
TRENDS IN NEUROSCIENCES, 1998, 21 (12) :529-537
[2]   THE EFFECT OF MODIFYING POTASSIUM CONCENTRATION ON THE INHIBITION OF MYOCARDIAL NA+-K+-ATPASE BY 2 CLASS-IB ANTIARRHYTHMIC DRUGS - LIDOCAINE AND TOCAINIDE [J].
ALMOTREFI, AA ;
DZIMIRI, N .
GENERAL PHARMACOLOGY, 1991, 22 (06) :1097-1101
[3]   Serious complications related to regional anesthesia - Results of a prospective survey in France [J].
Auroy, Y ;
Narchi, P ;
Messiah, A ;
Litt, L ;
Rouvier, B ;
Samii, K .
ANESTHESIOLOGY, 1997, 87 (03) :479-486
[4]  
BANKER G, 1991, CULTURING NERVE CELL, P11
[5]  
BIAN JH, 1991, J BIOL CHEM, V266, P8801
[6]   MOLECULAR MECHANISMS OF LOCAL-ANESTHESIA - A REVIEW [J].
BUTTERWORTH, JF ;
STRICHARTZ, GR .
ANESTHESIOLOGY, 1990, 72 (04) :711-734
[7]  
David G, 2000, J NEUROSCI, V20, P7290
[8]  
Gold MS, 1998, J PHARMACOL EXP THER, V285, P413
[9]  
Grinnell F, 1978, Int Rev Cytol, V53, P65
[10]   SODIUM-CHANNEL BLOCKADE REDUCES HYPOXIC SODIUM LOADING AND SODIUM-DEPENDENT CALCIUM LOADING [J].
HAIGNEY, MCP ;
LAKATTA, EG ;
STERN, MD ;
SILVERMAN, HS .
CIRCULATION, 1994, 90 (01) :391-399