title: Bryonolic Acid, a Triterpenoid, Protect Against N-methyl-d-Aspartate-Induced Neurotoxicity in PC12 Cells.
abstract: Calcium overload is considered to be one of the mechanisms of cerebral ischemia. Ca(2+) influx and Ca(2+)/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. Ca(2+) 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 Ca(2+) 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 Ca(2+) influx and regulating gene expression in the Ca(2+)-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.
authors.affiliation: College of Pharmacy, Fujian University of Traditional Chinese Medicine, Minhou Shangjie, Fuzhou 350122, Fujian, China. qjhfjtcm@sina.com.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Que
authors.affiliation: College of Pharmacy, Fujian University of Traditional Chinese Medicine, Minhou Shangjie, Fuzhou 350122, Fujian, China. AS2006041098@163.com.
authors.forename: Miao
authors.initials: M
authors.lastname: Ye
authors.affiliation: College of Pharmacy, Fujian University of Traditional Chinese Medicine, Minhou Shangjie, Fuzhou 350122, Fujian, China. zyqfj@hotmail.com.
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Pharmacy, Fujian University of Traditional Chinese Medicine, Minhou Shangjie, Fuzhou 350122, Fujian, China. xwfjtcm@126.com.
authors.forename: Wen
authors.initials: W
authors.lastname: Xu
authors.affiliation: College of Pharmacy, Fujian University of Traditional Chinese Medicine, Minhou Shangjie, Fuzhou 350122, Fujian, China. lihuang3413@gmail.com.
authors.forename: Huang
authors.initials: H
authors.lastname: Li
authors.affiliation: College of Pharmacy, Fujian University of Traditional Chinese Medicine, Minhou Shangjie, Fuzhou 350122, Fujian, China. xwfjlab@163.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Xu
authors.affiliation: College of Pharmacy, Fujian University of Traditional Chinese Medicine, Minhou Shangjie, Fuzhou 350122, Fujian, China. chukedan@gmail.com.
authors.forename: Kedan
authors.initials: K
authors.lastname: Chu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002545:Brain Ischemia / Q000139:chemically induced / Q000188:drug therapy*; D002118:Calcium / Q000378:metabolism; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000096:biosynthesis; D002470:Cell Survival / Q000187:drug effects; D017362:Cyclic AMP Response Element-Binding Protein / Q000096:biosynthesis; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D016202:N-Methylaspartate / Q000633:toxicity; D018696:Neuroprotective Agents / Q000008:administration & dosage* / Q000737:chemistry; D016716:PC12 Cells; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D014315:Triterpenes / Q000008:administration & dosage* / Q000737:chemistry; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis
chemical_list: C494828:Bax protein, rat; D017362:Cyclic AMP Response Element-Binding Protein; D018696:Neuroprotective Agents; D019253:Proto-Oncogene Proteins c-bcl-2; D014315:Triterpenes; D051028:bcl-2-Associated X Protein; D016202:N-Methylaspartate; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; C074984:bryonolic acid; D002118:Calcium
keywords: Bax; Bcl-2; Ca2+; PC12 cells; bryonolic acid; p-CREB; p-CaMKII