Fitoterapia 78 (2007) 235 237 www.elsevier.

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Short report

Effect of Azadirachta indica leaf methanol extracts on stem cell reproduction

M. Teresa Gonzlez-Garza a,, Margarita Codinach a,b , Citlali Alcaraz a , Jorge Moreno-Cuevas a , Pilar Carranza-Rosales c , Delia E. Cruz-Vega c
a

Divisin de Ciencias de la Salud, Centro de Investigacin y Extensin en Ciencias de la Salud, ITESM, C.P, 64710, Monterrey, Mexico b Facultat de Ciencies, Universitat Autonoma de Barcelona, 08193 Bellaterra Barcelona, Spain c Centro de Investigacin Biomdica del Noreste IMSS, C.P. 64710, Monterrey, Mexico Received 1 December 2005; accepted 27 November 2006 Available online 6 February 2007

Abstract Methanol extracts of Azadirachta indica leaves at concentration from 0.1 to 40 g/ml showed in vitro an stimulatory activity in stem cell reproduction. These results suggest that the effect of methanol leaf extracts on stem cell reproduction could be of benefit to improve health. 2007 Elsevier B.V. All rights reserved.
Keywords: Azadirachta indica; Stem cells; Reproduction

1. Plant Azadirachta indica leaves (Meliaceae), collected in October at a private nursery at San Nicolas, Nuevo Len, Mexico, were identified in the Herbarium of the Facultad de Ciencias Biolgicas de la UANL where a specimen (voucher no 02119) is deposited. 2. Uses in traditional medicine A. indica is a tree used worldwide in traditional medicine. The tree is considered a village pharmacy because its every part has an application in curing human diseases. Its constituents have been demonstrated to exhibit immunomodulatory, antiinflammatory, antihyperglycemic, antiulcer, antimalarial, antifungal, antibacterial, antiviral, antioxidant, antimutagenic and anticarcinogenic properties [1,2].

Corresponding author. Tel./fax: +52 81 8333 1121. E-mail address: mtgonzalezgarza@itesm.mx (M. Teresa Gonzlez-Garza). 0367-326X/$ - see front matter 2007 Elsevier B.V. All rights reserved. doi:10.1016/j.fitote.2006.11.016

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M. Teresa Gonzlez-Garza et al. / Fitoterapia 78 (2007) 235237

3. Previously isolated constituents Triterpenoids, limonoides, flavanones, tetranortriterpenoid and cathechins [37]. 4. Tested material Leaves were Soxhlet extracted with MeOH and concentrated in vacuo to give an extract (yield: 2.8%). The extract was dissolved in 10% DMSO in phosphate buffered saline for the experiments. 5. Studied activity Stem cell reproduction inducer. 6. Animals Two-month-old male Sprague Dawley rats were used. Animals were maintained in polypropylene cages with food (Lab. Diet. 5001, USA) and water ad libitum under controlled conditions of light, humidity and temperature. The animals used in the experimental protocol were treated ethically according to the guidelines of the World Health Organization. 7. Used cells Stem cells were obtained from bone marrow of adult rats. Cells recovered were incubated in Petri dishes with DMEM-F12 media supplemented with 10% fetal bovine serum (FBS) at 37 C with 5% CO2 in air. The cells were incubated with various concentrations of extract (0.1 to 250 g/ml) in 96-well microtiter plates with DMEM-F12 media supplemented with 10% FBS, at 37 C with 5% CO2 in air. Cell number was determined by soluble tetrazolium salt assay (Promega CellTiter 96 AQueous) after 72 h of incubation. Control cells were incubated with DMSO in phosphate buffer saline at the same concentration present in the samples. 8. Results A. indica leaf methanol extract between 0.1 and 40 g/ml gave an increase in cell number directly proportional to extract concentration. Concentration between 60 and 250 g/ml gave similar results in that the cell population doubled. Higher concentrations showed a slight decrease in the final population achieved compared with lower concentrations (Fig. 1).

Fig. 1. Effect of A. indica leaf methanol extract (0 to 250 g/ml) on stem cell reproduction after 72 h of cell incubation.

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9. Conclusions The results suggest that a benefit of A. indica leaf methanol extract administration could be an increase in stem cells. It is known that stem cells can repair damaged tissue [8,9]. The increase in multipotent cell numbers in the body could be responsible, at least in part, for improved general health in subjects using A. indica leaf methanol extract. References
[1] [2] [3] [4] [5] [6] [7] [8] [9] Subapriya R, Nagini S. Curr Med Chem Anti-Canc Agents 2005;5:149. Singh UP, Singh DP. J Herb Pharmacother 2002;2:13. Siddiqui BS, Afshan F, Gulzar T, Hanif M. Phytochemistry 2004;65:2363. Cohen E, Quistad GB, y Casida JH. Life Sci 1996;13:1075. Nakahara K, Roy MK, Ono H, Maeda I, Ohnishi-Kameyama M, Yoshida M, et al. J Agric Food Chem 2003;51:6456. Siddiqui S, Faizi S, Siddiqui BS, Ghiasuddin. J Nat Prod 1992;55:303. Luo, XD, Wu, SH, Ma, YB, Wu, DG. 2000;71:668. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Science 1999;284:143. Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, et al. Nature 2002;418:41.