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Estrogen activity of Silkworm (Bombyx mori) Pupa water extract and its fractions

  • Ryu, Jae-Sung (Department of Biological Science, College of Natural Sciences, Wonkwang University) ;
  • Jo, Gyeong-Jong (Professional Graduate School of Oriental Medicine and Traditional Oriental Medicine Institute, Wonkwang University) ;
  • Jin, Jung-Woo (Department of Biological Science, College of Natural Sciences, Wonkwang University) ;
  • Yang, Hyo-Jung (Department of Biological Science, College of Natural Sciences, Wonkwang University) ;
  • Park, Yong-Il (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea) ;
  • Na, Ye-Seul (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea) ;
  • Nam, Kyung-Su (Bogo F&D) ;
  • Keum, Kyung-Soo (Professional Graduate School of Oriental Medicine and Traditional Oriental Medicine Institute, Wonkwang University) ;
  • Choo, Young-Kug (Department of Biological Science, College of Natural Sciences, Wonkwang University)
  • Published : 2008.09.30

Abstract

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), $40^{\circ}C$, $60^{\circ}C$, $80^{\circ}C$, and $100^{\circ}C$ 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 ${\alpha}\;(ER{\alpha})$ and $ER{\beta}$, 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 $ER{\alpha}$ ($IC_{50}$, 1.76 ug/ml) and $ER{\beta}$ ($IC_{50}$, 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 ($H^+$) 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 ($Cl^-$) 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} ($IC_{50}$, 1.44 ug/ml) and $ER{\beta}$ ($IC_{50}$, 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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References

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