Neuroprotective Effect of Ginkgolide B on Bupivacaine-Induced Apoptosis in SH-SY5Y Cells

Le Li,Shi-yuan Xu,Qing-guo Zhang,Chi-wai Cheung,Xian-jie Wen,Ting Zheng,Shu-qin Zhou,Lu-ying Lai

doi:10.1155/2013/159864

Abstract

Local anesthetics are used routinely and effectively. However, many are also known to activate neurotoxic pathways. We tested the neuroprotective efficacy of ginkgolide B (GB), an active component of Ginkgo biloba, against ROS-mediated neurotoxicity caused by the local anesthetic bupivacaine. SH-SY5Y cells were treated with different concentrations of bupivacaine alone or following preincubation with GB. Pretreatment with GB increased SH-SY5Y cell viability and attenuated intracellular ROS accumulation, apoptosis, mitochondrial dysfunction, and ER stress. GB suppressed bupivacaine-induced mitochondrial depolarization and mitochondria complex I and III inhibition and increased cleaved caspase-3 and Htra2 expression, which was strongly indicative of activation of mitochondria-dependent apoptosis with concomitantly enhanced expressions of Grp78, caspase-12 mRNA, protein, and ER stress. GB also improved ultrastructural changes indicative of mitochondrial and ER damage induced by bupivacaine. These results implicate bupivacaine-induced ROS-dependent mitochondria, ER dysfunction, and apoptosis, which can be attenuated by GB through its antioxidant property.

Highlights

Local anesthetics are among the most common clinical drugs and are generally regarded as safe [1, 2]
The effect of bupivacaine on the viability of SH-SY5Y neuroblastoma cells was first examined using the by MTT assay
Bupivacaine (1, 1.5, 2 mmol/L) significantly reduced viable cell number compared to controls (Figure 1(a))

Summary

Introduction

Local anesthetics are among the most common clinical drugs and are generally regarded as safe [1, 2] They have been shown to be neurotoxic even at normal clinical dose [3, 4]. Intrathecally administered local anesthetics induced cell swelling, atrophy, edema, axonal degeneration, and the appearance of myelin ovoids as well as macrophage infiltration [7]. These morphological signs of degeneration indicate that local anesthetics can initiate a complex cascade of direct cytotoxic and ensuing inflammatory responses, the molecular mechanisms of local anesthetic toxicity are still largely unknown. Mitochondrial and ER damage associated with ROS overproduction may act synergistically to evoke cell death in response to bupivacaine or other structurally related local anesthetics

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