Bryonolic Acid, a Triterpenoid, Protect Against N-methyl-d-Aspartate-Induced Neurotoxicity in PC12 Cells.

Jinhua Que,Huang Li,Miao Ye,Wen Xu,Kedan Chu,Wei Xu,Yuqin Zhang

doi:10.3390/molecules21040418

Jinhua Que, Huang Li + Show 5 more

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https://doi.org/10.3390/molecules21040418

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Abstract

Calcium overload is considered to be one of the mechanisms of cerebral ischemia. Ca2+ influx and Ca2+/calmodulin-dependent protein kinase II (CaMKII) and cAMP response element-binding protein (CREB) phosphorylation are considered to be involved in N-Methyl-d-aspartate (NMDA)-induced apoptosis process. This study investigated the neuroprotective effects of bryonolic acid (BA) in an NMDA-induced rat adrenal pheochromocytoma cell line (PC12) cells and the potential mechanism. PC12 was treated by NMDA to establish an excitotoxicity model. BA (110,100 and 1000 μM final concentration) was added to the medium 24 h prior to the addition of NMDA. Subsequently, a methyl thiazolyl tetrazolium (MTT) assay and a lactate dehydrogenase (LDH) release were performed. Ca2+ concentration was demonstrated using a scanning-dual wavelength fluorimetric method. In addition, protein and mRNA levels were determined via Western blot and real-time PCR. In the presence of BA, MTT assay and LDH assay showed that more cells were viable in comparison with the NMDA group. Moreover, the concentration of Ca2+ decreased with the addition of BA in culture. Furthermore, BA could upregulate protein expressions of Bcl-2, p-CREB, and p-CaMKII and downregulate protein expression of Bax. The mRNA results showed that the pattern of mRNA expression were similar to their respective protein levels. All these results indicate that BA protected PC12 cells against NMDA-induced apoptosis by inhibiting Ca2+ influx and regulating gene expression in the Ca2+-CaMKII-CREB signal pathway. Therefore, the present study supports the notion that BA may be a promising neuroprotective agent for the treatment of cerebral ischemia disease.

Highlights

IntroductionIschemic stroke is by far the most common kind of stroke that accounts for about 88% of all strokes
Ischemic stroke is by far the most common kind of stroke that accounts for about 88% of all strokes.It is the major cause of disability and the third cause of death across the world [1]
NMDA is a kind of excitatory amino acid, which acts on N-methyl-D-aspartate receptors (NMDAR) of postsynaptic neuron, activates calcium channel controlled by receptor, and receptors (NMDAR) of postsynaptic neuron, activates calcium channel controlled by receptor, and results in calcium overload

Summary

Introduction

Ischemic stroke is by far the most common kind of stroke that accounts for about 88% of all strokes. It is the major cause of disability and the third cause of death across the world [1]. The mechanisms involved in ischemic stroke is complex and not fully understood, and excitotoxicity, calcium overload, oxidative stress, inflammation, and apoptosis [2,3,4,5] are involved. It was proved that cerebral ischemic injury was related to massive influx of extracellular Ca2+ [6,7]. Calcium overload is regarded as the final common pathway of the action mechanisms leading to neuronal death [8,9,10].

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