脂肪乳剂救治布比卡因致大鼠惊厥对海马神经元凋亡的影响

Effect of Lipid Emulsion on the Apoptosis of Hippocampal Neurons Induced by Bupivacaine Induced Seizures in Rats

目的研究脂肪乳剂救治布比卡因致惊厥大鼠对海马神经元凋亡的影响。方法 45只健康雄性SD大鼠,随机分3组(n=15):空白对照组(Control),布比卡因组(BPV+NS),脂肪乳剂处理组(BPV+LE)。Control组大鼠先给予生理盐水0.13 m L·(kg·min)-1匀速泵注9 min,15 s,再以3 m L·(kg·min)-1的速率泵注5 min制备对照模型;(BPV+NS)组给予0.75%的布比卡因1 mg·(kg·min)-1至大鼠惊厥,立即给予生理盐水3 m L·(kg·min)-1,5min处理;(BPV+LE)组给予布比卡因1 mg·(kg·min)-1至大鼠惊厥,立即给予肪乳剂3m L·(kg·min)-1,5min处理。记录大鼠惊厥时布比卡因的用量及惊厥持续时间。造模后24h取标本,Western blot检测大鼠海马组织cleaved caspase-3蛋白的表达,TUNEL法检测海马神经元凋亡率,电镜观察海马神经元超微结构。结果 (BPV+NS)组与(BPV+LE)组大鼠惊厥时布比卡因的用量及惊厥开始时间差异无统计学意义(P>0.05),而(BPV+LE)组大鼠惊厥持续时间(376.7±123.5)s短于(BPV+NS)组(592.0±223.3)s(P<0.01);造模后24h各组大鼠海马组织cleaved caspase-3蛋白表达量差异无统计学意义,海马神经元凋亡率差异无统计学意义(P均>0.05);电镜观察(BPV+NS)组和(BPV+LE)组海马神经元线粒体嵴溶解断裂、内质网水肿及突触融合现象增多。结论脂肪乳剂能改善布比卡因致大鼠中枢神经毒性症状,对大鼠布比卡因惊厥后24h海马神经元凋亡情况无明显影响。

Objective To investigate the effect of the apoptosis on lipid emulsion rescue hippocampal neurons in rats of central nervous toxicity induced by bupivacaine in rats. Methods 45 healthy male SD rats were randomly divided into three groups（n=15）：control group（Control）,bupivacaine＋normal saline group （BPV＋NS）and bupivacaine ＋ lipid emulsion group（BPV＋LE）. Rats in the control group were given normal saline at the rate of 0.13 mL·（kg·min）-1 for 9 min,3mL·（kg·min）-1 for 5 min successively to built control model. BPV＋NS group and BPV＋LE group rats were given 0.75% bupivacaine 1 mg·（kg·min）-1 to prepare bupivacaine convulsion model rat,then treated with normal saline or lipid emulsion at rate of 3 mL·（kg· min）-1 for 5 min,and the dosage of bupivacaine and the duration of convulsion were recorded when the rats convulsion. The expression of cleaved caspase-3 protein in the hippocampus was detected by Western blot,the neurons apoptosis in hippocampal were detected by TUNEL,and the ultrastructure of hippocampal neurons was observed by electron microscope after 24 hours of modeling. Results There was no statistical difference in the dosage of bupivacaine between BPV＋NS group and BPV＋LE group,the convulsion duration of the BPV＋LE group（376.7 ±123.5）s was significantly shorter than BPV＋NS group（592.0 ±223.3）s（P〈0.01）. There were no statistical differences in the expression of cleaved caspase-3 protein in the hippocampus and the neurons apoptosis of hippocampal were detected by TUNEL among three groups. Electron microscope observation showed that BPV＋NS compared with control group and BPV＋LE group,there were increase in mitochondrial crest disintegration,endoplasmic reticulum edema and synaptic fusion of hippocampal neuronal. Conclusion Lipid emulsion can significantly improve the syndrome of the central nervous system toxicity induced by bupivacaine in rats, but there is no significant effect on hippocampal neuronal apoptosis after 24 hours of rats convulsion. This study provides a reference for investigation the mechanism of lipid emulsions in treatment of central nervous system toxicity induced by bupivacaine .