Elsevier

Journal of Ethnopharmacology

Volume 127, Issue 2, 3 February 2010, Pages 229-234
Journal of Ethnopharmacology

Osteoprotective effect of extract from Achyranthes bidentata in ovariectomized rats

https://doi.org/10.1016/j.jep.2009.11.016Get rights and content

Abstract

Aim of the study

A Chinese herbal drug, root of Achyranthes bidentata showed a potent inhibitory activity on bone resorption induced by parathyroid hormone (PTH) in a bone organ culture using neonatal mouse parietal bones. The present study is to clarify the fractions responsible for the activity and further explore the osteoprotective effect of the fraction in vivo.

Materials and methods

The hexane, ethyl acetate (EtOAc), n-butanol (n-BuOH) and water soluble fractions of methanol extract of the root of Achyranthes bidentata were prepared and screened for their anti-bone resorption activity using the bone organ culture system. The n-BuOH soluble fraction was further administered orally at doses of 25, 50 and 100 mg/(kg day) to ovariectomized (OVX) rats. The analyses of the rat body weight, serum estradiol (E2), total cholesterol and triglyceride levels, uteri weight and measurement of bone mineral density (BMD) were conducted.

Results

The EtOAc and n-BuOH fractions showed the most potent inhibitory activity on PTH-induced bone resorption. Further research using OVX rat model revealed that the n-BuOH fraction significantly prevented BMD loss due to OVX operation. While, the uteri weight and serum estradiol (E2), total cholesterol and triglyceride levels displayed no differences compared with those of control group (OVX rats), suggesting the n-BuOH fraction should have no estrogen-like side effects.

Conclusions

The results reveal that the n-BuOH soluble fraction of the root of Achyranthes bidentata is effective at preventing bone loss in OVX rats and has a great potential as an alternative tool for the treatment of osteoporosis.

Graphical abstract

The n-butanol soluble fraction of methanol extract from root of Achyranthes bidentata displayed a remarkable osteoprotective effect in ovariectomized rats.

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Introduction

Osteoporosis is a disease characterized by low bone mass and structural deterioration of bone tissue, leading to bone fragility and an increased susceptibility to fractures (Epstein, 2006). It is widely recognized as a major public health problem, affecting both female and male. Most cases of osteoporosis occur in postmenopausal women due to the dramatic estrogen withdrawal associated with menopause. In America, it threats an estimated 44 million people, or 55 percent of the people of 50 years of age and older (Buencamino et al., 2009). In Europe, osteoporosis related fractures accounts for more disability-adjusted life years (DALYs) lost than common cancers except for lung cancer (Johnell and Kanis, 2006). In China, it is estimated that over 90 million people are suffering from osteoporosis (Hu et al., 2008). Currently, hormone replacement therapy (HRT) is an established method in the prevention of fractures and bone loss for postmenopausal women, however, it demonstrates an increased risk of breast cancer, stroke, and heart attacks, which are generally called estrogen-like side effects (Canderelli et al., 2007). Therefore, the compliance of HRT decreases and many doctors and patients look for alternatives (Wuttke et al., 2008). Recent results provide a potential that the use of alternative strategies for the management of osteoporosis sourced from some medicinal plants and natural products to avoid the increased significant risks might be achieved (Penolazzi et al., 2008, Doyle et al., 2009).

In traditional Chinese medicine, there are a number of crude drugs that possess functions to ease the joints, strengthen bones and muscles, and have been used for treatment of bone injuries and bone related diseases. In a research project aimed at finding active extracts or compounds from natural resources with potential of osteoprotective effect, we have screened a number of crude drug extracts for their inhibitory activity on bone resorption induced by parathyroid hormone (PTH) in a bone organ culture system (Li et al., 1996). Among the extracts, a methanol extract from the root of Achyranthes bidentata Blume (Amaranthaceae) showed the most potent inhibitory activity at the concentration of 44 μg/mL. Achyranthes bidentata, as a very important herbal drug listed in Chinese Pharmacopoeia (China Pharmacopoeia Committee, 2005) possesses medical functions of invigoration of liver–kidney and strengthening of muscle–bones, and has been used to treat bone related diseases for a long time. It is reported that the polysaccharides from this medicinal plant have oxidative stress protective effect (Xue et al., 2009), anti-HIV activity (Peng et al., 2008) and inhibit non-enzymatic glycation in a d-galactose induced mouse aging model (Deng et al., 2003). The saponins from the plant showed adjuvant effect on specific antibody and cellular response to ovalbumin (Sun, 2006). Two compounds, ecdysterone and daucosterol isolated from this plant markedly stimulated proliferation of osteoblast-like UMR106 cells (Li et al., 2001). To clarify the active fractions responsible for the anti-bone resorption activity and further explore the osteoprotective effect in vivo, a detailed investigation on Achyranthes bidentata was conducted. In the present study, the hexane, ethyl acetate (EtOAc), n-butanol (n-BuOH) and water soluble fractions of methanol extract of Achyranthes bidentata were prepared and tested for their anti-bone resorption activity. The in vivo effect of the n-BuOH fraction on uterine weight, serum estradiol (E2), total cholesterol and triglyceride levels, and the maintenance of bone mineral density (BMD) of lumbar spine (L2–4) were further investigated in ovariectomized (OVX) rat.

Section snippets

Materials

The roots of Achyranthes bidentata Blume (Amaranthaceae) were purchased from the Department of Medicinal Materials, Jiangsu Province Hospital of Traditional Chinese Medicine, and identified by Prof. Z.N. Gong (School of Life Science, Nanjing Normal University). Voucher specimen has been deposited in the School of Chemistry and Chemical Engineering of Nanjing University (No. TCM060112).

The chemicals and reagents were obtained from the following sources: Ham's F-12 medium from Nissui

Inhibitory activity of the fractions on PTH-induced bone resorption

Because the methanol extract of Achyranthes bidentata showed the strongest inhibitory activity on PTH-induced bone resorption (Li et al., 1996), the methanol extract was further fractionationed to hexane, EtOAc, n-BuOH and water soluble fractions. The inhibitory activity on bone resorption of the four fractions was examined. As shown in Table 1, hexane fraction displayed no inhibitory activity and water fraction showed a weak activity only at the highest concentration of 440 μg/mL. However,

Discussion

Although Achyranthes bidentata has been a ‘traditional medicine’ to strengthen the bone for many years, to date, no in-depth study has been carried out to determine the action. In the present study, we have evaluated the effects of this herbal plant on osteoporosis using neonatal mouse bone organ culture system and OVX rats.

Parathyroid hormone (PTH), a polypeptide secreted by the parathyroid glands is responsible for relatively small changes in serum Ca2+. Together with the vitamin D endocrine

Conclusions

The present study indicated that the n-BuOH fraction of Achyranthes bidentata displayed osteoprotective effect on bone loss in OVX rats, and has a great potential as a therapeutic tool against osteoporosis. Although the inhibitory effect on bone resorption should closely relate to the preventive action on the decrease of BMD, the association of the action with female hormone is still unclear. To elucidate the molecular mechanisms involved in the action, further research would be essential, and

Acknowledgements

This work was supported by National Natural Science Fund for Creative Research Groups of China (20521503) and 973 Program (2007CB714504).

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