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KMID : 1200820080080030228
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Oriental Pharmacy and Experimental Medicine
2008 Volume.8 No. 3 p.228 ~ p.235
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Estrogen activity of Silkworm (Bombyx mori) Pupa water extract and its fractions
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Ryu Jae-Sung
Jo Gyeong-Jong
Jin Jung-Woo
Yang Hyo-Jung
Park Yong-Il
Na Ye-Seul
Nam Kyung-Su
Keum Kyung-Soo
Choo Young-Kug
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Abstract
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This study was conducted to evaluate the estrogen activity of silkworm (Bombyx mori) pupa extracts and their fractions. Powdered samples of freeze-dried silkworm pupa were extracted at room temperature (RT), , , , and in water (D.W), chloroform, ethyl acetate, and methanol for 6h and then filtered (0.45 um). The extracts were then freeze-dried. The estrogenic activity of these extracts was then investigated by competition binding assays using estrogen receptor and , and by evaluating their effects on the proliferation of the human breast cancer cell line, MCF-7. Among the extracts evaluated, water extracts prepared at RT showed the highest binding affinity to (, 1.76 ug/ml) and (, 0.07 ug/ml). In addition, MCF-7 cells that were treated with 62.5 ug/ml of the RT extract showed the greatest increase in proliferation (2-fold; 1291.79%) when compared to control cells (659.82%). Next, the water extract that was prepared at RT (sample 1) was dissolved in D.W. and further fractionated using a Dowex 50W - 8X () column. The flow-through and wash were then pooled together and freeze-dried (sample 2). The bound materials were then eluted with 20 mM NaCl, after which they were applied to a Dowex 1X2 - 200 () column and washed with D.W. to remove the sodium ions. The eluants were then freeze-dried (sample 3). Of these fractions, sample 2 showed the highest binding affinity to ER{\alpha} (, 1.44 ug/ml) and (, 1.18 ug/ml). In addition, MCF-7 cells that were treated with sample 2 (15.6 ug/ml) showed the largest increase in growth (1159.39%) when compared to control cells (525.26%). Taken together, these results suggest that the fraction of the RT water extract of silkworm pupa referred to as sample 2 may be useful as a phytoestrogen.
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KEYWORD
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Silkworm pupa water extract, Estrogen, Estrogen receptor, MCF-7 cells
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