Send Orders for Reprints to reprints@benthamscience.net Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry, 2014, 13, 45-55 45 Beta Caryophyllene and Caryophyllene Oxide, Isolated from Aegle Marmelos, as the Potent Anti-inflammatory Agents against Lymphoma and Neuroblastoma Cells Soumyadeep Sain 1 , Pravin K. Naoghare 1 , S. Saravana Devi 1,* , Atul Daiwile 1 , K. Krishnamurthi 1 , P. Arrigo 2 and T. Chakrabarti 1 1 Environmental Health Division, National Environmental Engineering Research Institute (NEERI), CSIR, Nagpur, India; 2 CNR Institute for Macromolecular Studies, Genoa, Italy Abstract: Aegle marmelos (Indian Bael) is a tree which belongs to the family of Rutaceae. It holds a prominent position in both Indian medicine and Indian culture. We have screened various fractions of Aegle marmelos extracts for their anti- cancer properties using in vitro cell models. Gas chromatography-Mass spectrometry (GC-MS) was employed to analyze the biomolecules present in the Aegle marmelos extract. Jurkat and human neuroblastoma (IMR-32) cells were treated with different concentrations of the fractionated Aegle marmelos extracts. Flow cytometric analysis revealed that optimal concentration (50g/ml) of beta caryophyllene and caryophyllene oxide fractions of Aegle marmelos extract can induce apoptosis in Jurkat cell line. cDNA expression profiling of pro-apoptotic and anti-apoptotic genes was carried out using real time PCR (RT-PCR). Down-regulation of anti-apoptotic genes (bcl-2, mdm2, cox2 and cmyb) and up-regulation of pro-apoptotic genes (bax, bak1, caspase-8, caspase-9 and ATM) in Jurkat and IMR-32 cells treated with the beta caryo- phyllene and caryophyllene oxide fractions of Aegle marmelos extract revealed the insights of the downstream apoptotic mechanism. Furthermore, in-silico approach was employed to understand the upstream target involved in the induction of apoptosis by the beta caryophyllene and caryophyllene oxide fractions of Aegle marmelos extract. Herein, we report that beta caryophyllene and caryophyllene oxide isolated from Aegle marmelos can act as potent anti-inflammatory agents and modulators of a newly established therapeutic target, 15-lipoxygenase (15-LOX). Beta caryophyllene and caryophyllene oxide can induce apoptosis in lymphoma and neuroblastoma cells via modulation of 15-LOX (up-stream target) followed by the down-regulation of anti-apoptotic and up-regulation of pro-apoptotic genes. Keywords: Apoptotic genes, caryophyllene, 15-lipoxygenase, lymphoma, neuroblastoma cells. INTRODUCTION Natural products have played an important role in the field of medicine which has been acknowledged [1]. Since the discovery of penicillin there has been a plethora of com- pounds originating from natural sources that showed anti- diabetic, anti-bacterial, anti-fungal or anti-inflammatory properties [2, 3]. The interest in natural products experienced a cessation since the advent of new synthetic drug analogs [4, 5]. Recently, the study of natural products has gained considerable importance in the field of medicine due to the significant side effects of synthetic drug analogs [5-7]. However, working with natural products has inherent drawbacks as the extract prepared from these products often represents a mixture of compounds and the active compound cannot be readily identified from this mixed pool [8, 9]. To accomplish this, time consuming process of purification ac- companied by screening of each fraction has to be carried out until we get the active compound. However, with the recent developments in analytical methods like liquid *Address correspondence to this author at the Environmental Health Division, National Environmental Engineering Research Institute (NEERI), CSIR, Nagpur, India; Tel:/Fax: + 91-712-2249961; E-mail: ss_devi@neeri.res.in chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-mass spectrometry (GC-MS), it is possible to rapidly identify the components of the crude ex- tract [8]. In addition, in silico modeling can be used as a powerful tool to screen the active bio-molecules from the mixture of compounds with pharmacological potentials [10- 12]. Thus, the application of advanced analytical technique to identify the components of the crude extracts followed by in-silico screening of this identified molecule against the cellular targets can lead to rapid identification of the active components in the crude extract together with the up-stream therapeutic targets of the molecular pathway. Aegle marmelos is a deciduous tree belonging to the fam- ily of rutaceae, which is commonly found all over India. It is important in contemporary Indian medicine as well as in the culture of the people of India. It is very rich in bioactive compounds that show activity against various human ail- ments [13]. Different compounds have been identified by chemical analysis, of which many showed biological activity against human diseases [14]. In this study, we have screened different fractions of Aegle marmelos leaves for their anti-inflammatory proper- ties. Beta caryophyllene and caryophyllene oxides isolated 1875-614X/14 $58.00+.00 © 2014 Bentham Science Publishers