Author's personal copy Tianma modulates proteins with various neuro-regenerative modalities in differentiated human neuronal SH-SY5Y cells Umamaheswari Ramachandran a,b , Arulmani Manavalan a,b , Husvinee Sundaramurthi a , Siu Kwan Sze a , Zhi Wei Feng a , Jiang-Miao Hu c , Klaus Heese a,b,⇑ a School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore b Institute of Advanced Studies, Nanyang Technological University, 60 Nanyang View, Singapore 639673, Singapore c Kunming Institute of Botany, Chinese Academy of Science, Kunming, Yunnan 650204, People’s Republic of China article info Article history: Received 10 November 2011 Received in revised form 29 February 2012 Accepted 19 March 2012 Available online 28 March 2012 Keywords: Tianma Neuron Herb TCM iTRAQ Neurodegeneration abstract Tianma (Rhizoma gastrodiae) is the dried rhizome of the plant Gastrodia elata Blume (Orchidaceae family). As a medicinal herb in traditional Chinese medicine (TCM) its functions are to control convulsions, pain, headache, dizziness, vertigo, seizure, epilepsy and others. In addition, tianma is frequently used for the treatment of neurodegenerative disorders though the mechanism of action is widely unknown. Accord- ingly, this study was designed to examine the effects of tianma on the proteome metabolism in differen- tiated human neuronal SH-SY5Y cells to explore its specific effects on neuronal signaling pathways. Using an iTRAQ (isobaric tags for relative and absolute quantitation)-based proteomics research approach, we identified 2390 modulated proteins, out of which 406 were found to be altered by tianma in differenti- ated human neuronal SH-SY5Y cells. Based on the observed data, we hypothesize that tianma promotes neuro-regenerative signaling cascades by controlling chaperone/proteasomal degradation pathways (e.g. CALR, FKBP3/4, HSP70/90) and mobilizing neuro-protective genes (such as AIP5) as well as modulating other proteins (RTN1/4, NCAM, PACSIN2, and PDLIM1/5) with various regenerative modalities and capac- ities related to neuro-synaptic plasticity. Ó 2012 Elsevier Ltd. All rights reserved. 1. Introduction Despite the many cell- and gene-based therapeutic efforts made for the treatment of neurological disorders, such as Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD) and others, these diseases are still fatal and a continuous threat to our aging societies as they result in increasing loss of cognitive and physical functions. Since currently no effective therapy is in place for the treatment of such diseases, recent data predict that the number of people affected by AD and aging-related dementia will continue to grow at an epidemic pace (Both et al., 2011; Chiu and Rao, 2011; Heese et al., 2006; Karran et al., 2011). Increasing attention has thus led to enforced efforts in search of suitable novel medicinal herbs for therapeutic applications (Schachter, 2009; Sucher, 2006; Yuan and Lin, 2000), in particular for the treatment of dementia (Rafii et al., 2011). Orchids and their derivatives have been shown to improve neu- ral functions in clinical studies but the underlying mechanisms are largely unknown which severely hampered the more extensive application of such potential drugs as well as the potential industrial exploration of it (Bulpitt, 2005; Bulpitt et al., 2007; Hew and Yong, 2007). Gastrodia elata Blume, known as tianma in Chinese, is a plant of the orchidaceae family, the tuber of which can be dried and used as a medicinal material. In traditional Chinese medicine theory, tianma has the functions of pacifying the liver and calming wind. Besides, pharmacological studies indicate that tianma has the ef- fects of anti-convulsion, sedation, analgesia, memory improvement, anti-aging, improvement of microcirculation, lowering of blood pressure, improvement of blood vessel functions and anti-inflam- matory immunostimulation. In addition, it is often used for the treatment of cardio-, cerebro- and neurovascular diseases including migraine, depression and epilepsy (Bulpitt et al., 2007; Hew and Yong, 2007; Hsieh et al., 1997; Kim et al., 2003a,b; Kim et al., 2001; Ong et al., 2007). Previously, we demonstrated in vivo the po- tential neuro-protective action of tianma and its capacity to en- hance cognitive functions in mice (Mishra et al., 2011b). Recently, we have successfully applied the two dimensional (2D) liquid chromatography coupled with tandem mass spectrom- etry-based isobaric tag for relative and absolute quantification (2D-LC-MS/MS-iTRAQ) strategy in the area of neuro-degenerative diseases (Datta et al., 2010,2011). Here we used this proteomics approach in our in vitro human neural SH-SY5Y cell model for quantitative profiling of tianma-regulated genes. In quest of the 0197-0186/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.neuint.2012.03.012 ⇑ Corresponding author. Address: School of Biological Sciences, Nanyang Tech- nological University, 60 Nanyang Drive, Singapore 637551, Singapore. Tel.: +65 6316 2848; fax: +65 6791 3856. E-mail address: Klaus.Heese@rub.de (K. Heese). Neurochemistry International 60 (2012) 827–836 Contents lists available at SciVerse ScienceDirect Neurochemistry International journal homepage: www.elsevier.com/locate/nci