A water-soluble high molecular weight substance isolated from Hyuganatsu orange (Citrus tamurana), suspected to be a polysaccharide, inhibits rat osteoclast cell formation

Authors

  • Hiroko Hata Department of Obstetrics and Gynecology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake-Cho, Miyazaki City, Miyazaki 889-1692, Japan
  • Masatoshi Yamaguchi Department of Obstetrics and Gynecology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake-Cho, Miyazaki City, Miyazaki 889-1692, Japan
  • Hiroshi Sameshima Department of Obstetrics and Gynecology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake-Cho, Miyazaki City, Miyazaki 889-1692, Japan
  • Tsuyomu Ikenoue Department of Obstetrics and Gynecology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake-Cho, Miyazaki City, Miyazaki 889-1692, Japan
  • Junko Matsubara Ichimaru Phalcos Company, 318-1, Asagi, Motosu City, Gifu 501-0475, Japan
  • Makoto Tsuboi Ichimaru Phalcos Company, 318-1, Asagi, Motosu City, Gifu 501-0475, Japan
  • Takashi Tanaka Laboratory of Natural Product Chemistry, Nagasaki University Graduate School of Biomedical Sciences, 1-14 Bunkyo-cho, Nagasaki City, Nagasaki 852-8521, Japan

DOI:

https://doi.org/10.31989/ffhd.v5i6.182

Abstract

Background: Osteoporosis is detrimental to aged women’s health care. We previously reported that Hyuganatsu orange (Citrus Tamurana) contains active substances that inhibit osteoclast activities. Prior to conducting a human study, we sought to identify the biological active substance in the Hyuganatsu orange which suppresses osteoclast formation.

Methods: We isolated five fractions from a Hyuganatsu orange extract according to molecular weight. Each fraction was tested to determine its suppressive effect on the formation of osteoclasts in rats. We also used high-performance liquid chromatography (HPLC), infra-red (IR), and 1H and 13C NMR spectroscopy to evaluate its chemical structure. Data was recorded as mean ± standard error of the mean. The Mann-Whitney test was used, and a p-value of <.05 was considered statistically significant.

Results: The highest and lowest molecular weight fractions showed significant suppression activity on rat osteoclast formation (p < .05). The lowest molecular weight fraction was identified as hesperidin using thin layer chromatography. Additionally, IR absorption revealed that the highest molecular weight fraction was not a flavonoid. With regard to chemical structure, 1H and 13C NMR spectroscopy suggested that the highest molecular weight fraction had signals compatible with a polysaccharide such as galacturonic acid.

Conclusions: Hyuganatsu orange contains a biological active substance other than hesperidin that may be a polysaccharide and may suppress osteoclast formation.

Key Words: Citrus Tamurana, TRAP positive cell, Polysaccharides, Rat osteoclast cell, in vitro study

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Published

2015-06-01

Issue

Vol. 5 No. 6 (2015): June 2015

Section

Research Articles

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