The Effects Of Eurycoma Longifolia Extract And Eurycomanone On The Steroidogenesis In Mouse Leydig Cells
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Date
2018-07
Authors
Khurshid Ahmed, Nor Amira
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Eurycoma longifolia Jack (Tongkat Ali) is one of the most popular traditional
plants found in the tropical rainforest of Southeast Asia, especially in Malaysia and
Indonesia. Although E. longifolia is widely used as traditional medicine, little is
understood of the in vitro effect on Leydig cells; the testosterone-producing cells.
Therefore, the effect of a standardised E. longifolia extract (F2) and pure compound, eurycomanone (EN) on the productions of testosterone, estrogen and aromatase in
mouse Leydig cells were determined in this study, whereby the Formestane (FM) was used as a positive control. The effect of F2, EN and FM on the cell viability was first
determined using MTT assay. The cell cycle of the treated Leydig cells was then
subjected to Propidium iodide (PI) staining prior analysed by Flow Cytometry. The levels of testosterone, estrogen and aromatase in treated Leydig cells were then
determined by ELISA. Finally, the aromatase enzyme activity was screened using a high throughput inhibitor screening kit. The study found that the growth inhibitory effect increased in a dose-dependent manner in the Leydig cells treated with F2.
However, this phenomenon was not observed in EN- and FM-treated cells despite when higher concentrations were used. Cell cycle analysis revealed that F2-treated cells reduced viability due to the cell cycle arrest at S phase within 96 hours treatment. A significant S phase cell cycle arrest at 24 hours and 48 hours treatments could also be observed in EN-treated cells. However, the arrested cells were reduced at 72 hours and 96 hours treatments. Meanwhile, FM-treated cells did not show any significant
cell cycle arrest. The ELISA revealed that F2-, EN- and FM-treated Leydig cells
increased the level of testosterone may be due to the reduction of the conversion of
testosterone to estrogen.
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Keywords
The effects of eurycoma longifolia extract and eurycomanone , on the steroidogenesis in mouse leydig cells