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Molecules 2016, 21(4), 418; doi:10.3390/molecules21040418

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

College of Pharmacy, Fujian University of Traditional Chinese Medicine, Minhou Shangjie, Fuzhou 350122, Fujian, China
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Author to whom correspondence should be addressed.
Academic Editors: Vassilios Roussis and Efstathia Ioannou
Received: 9 January 2016 / Revised: 10 March 2016 / Accepted: 23 March 2016 / Published: 28 March 2016
(This article belongs to the Special Issue Triterpenes and Triterpenoids 2016)
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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. View Full-Text
Keywords: bryonolic acid; PC12 cells; Ca2+; Bcl-2; Bax; p-CaMKII; p-CREB bryonolic acid; PC12 cells; Ca2+; Bcl-2; Bax; p-CaMKII; p-CREB
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Que, J.; Ye, M.; Zhang, Y.; Xu, W.; Li, H.; Xu, W.; Chu, K. Bryonolic Acid, a Triterpenoid, Protect Against N-methyl-d-Aspartate-Induced Neurotoxicity in PC12 Cells. Molecules 2016, 21, 418.

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