Antiosteoporotic chemical constituents from Er-Xian Decoction, a traditional Chinese herbal formula.
OBJECTIVE
Er Xian Decoction (EXD), a traditional Chinese medicine formula, has long been used for the treatment of osteoporosis and menopausal syndrome in China. The present study was designed to investigate the antiosteoporotic constituents of EXD, and evaluate their antiosteoporotic effects in ovariectomized rats.


METHODS
Osteoblasts in neonatal calvaria cultures and osteoclasts derived from rat marrow cells were used to bioactivity-guided screen the active constituents. The proliferation of osteoblast was assayed by MTT methods. The activity of ALP and TRAP was measured by p- nitrophenyl sodium phosphate assay. The antiosteoporotic effects of icariin (1), anemarsaponin B II (8) and berberine (6) were verified by using OVX rats model. The bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry using the small animal scan mode. The undecalcified longitudinal proximal tibial metaphysical (PTM) sections were cut and stained for the bone histomorphometric analysis.


RESULTS
Bioactivity-guided fractionation has led to the successful isolation of antiosteoporotic constituents, i.e., icariin (1), icariside I (2), baohuoside I (3), mangiferin (4), neomangiferin (5), berberine (6), anemarsaponin B (7), anemarsaponin BII (8), anemarsaponin C (9), anemarrhenasaponin I (10), rubiadin-1-methyl ether (11) and obaculactone (12) from EXD. Further study showed that icariin (1), anemarsaponin BII (8) and berberine (6) increased the BMD in ovariectomized rats, and icariin (1) not only increased the bone formation, but also inhibited bone resorption; anemarsaponin BII (8) mainly increased bone formation and berberine (6) only inhibited the bone resorption in ovariectomized rats.


CONCLUSIONS
Our findings demonstrate that multiple ingredients are responsible for antiosteoporotic activity in traditional Chinese medicine formula Er-Xian decoction.
Qin
Luping
L
School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, PR China.
Han
Ting
T
Zhang
Qiaoyan
Q
Cao
Dapeng
D
Nian
Hua
H
Rahman
Khalid
K
Zheng
Hanchen
H
D015502:Absorptiometry, Photon; D000469:Alkaline Phosphatase / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D015519:Bone Density / Q000187:drug effects*; D001862:Bone Resorption / Q000517:prevention & control; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D005591:Chemical Fractionation; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000494:pharmacology; D005260:Female; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D010024:Osteoporosis / Q000188:drug therapy*; D010052:Ovariectomy; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar
D004365:Drugs, Chinese Herbal; C517980:er-xian decoction; D000469:Alkaline Phosphatase