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. 2005 Sep;2(3):353-61.
doi: 10.1093/ecam/neh101. Epub 2005 Jul 26.

The osteoprotective effect of Herba epimedii (HEP) extract in vivo and in vitro

Fang Xie  1 , Chun-Fu Wu, Wan-Ping Lai, Xu-Juan Yang, Pik-Yuan Cheung, Xin-Sheng Yao, Ping-Chung Leung, Man-Sau Wong
Affiliations

"VSports手机版" The osteoprotective effect of Herba epimedii (HEP) extract in vivo and in vitro

Fang Xie et al. Evid Based Complement Alternat Med. 2005 Sep.

Abstract

Herba epimedii (HEP) is one of the most frequently used herbs prescribed for treatment of osteoporosis in China. In the present study, the in vivo effects of HEP extract on bone metabolism were evaluated using 4-month-old ovariectomized (OVX) or sham-operated (Sham) female Sprague-Dawley rats orally administered with HEP extract (110 mg kgd), 17ss-estrogen (2 mg kgd) or its vehicle for 3 months. HEP extract significantly decreased urinary calcium excretion, suppressed serum alkaline phosphatase (ALP) activity and urinary deoxypyridinoline levels in OVX rats (P < 0. 05 versus vehicle-treated OVX rats). Histomorphometric analysis indicated that HEP extract could prevent OVX-induced bone loss by increasing tibial trabecular bone area and decreasing trabecular separation in OVX rats (P < 0. 05 versus vehicle-treated OVX group). The in vitro effects of HEP extract were also studied using rat osteoblast-like UMR 106 cells. HEP extract significantly stimulated cell proliferation in a dose-dependent manner (P < 0. 01 versus vehicle-treated) and increased ALP activity at 200 microgml (P < 0 VSports手机版. 01 versus vehicle-treated) in UMR 106 cells. It modulated osteoclastogenesis by increasing osteoprotegrin (OPG) mRNA and decreasing receptor activator of NF-kappaB ligand (RANKL) mRNA expression, resulting in a dose-dependent increase in OPG/RANKL mRNA ratio (P < 0. 01 versus vehicle-treated). Taken together, HEP treatment can effectively suppress the OVX-induced increase in bone turnover possibly by both an increase in osteoblastic activities and a decrease in osteoclastogenesis. The present study provides the evidence that HEP can be considered as a complementary and alternative medicine for treatment of post-menopausal osteoporosis. .

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Figures

<b>Figure 1</b>
Figure 1
Reverse-phase HPLC for the quantification of icarrin in HEP. The mobile phase was MeOH:H2O = 55:45 at 1 ml min−1. Peaks eluting at 18.5 min are icarrin.
<b>Figure 2</b>
Figure 2
Effect of HEP extract on body weight and uterine weight in OVX rats. (A) Body weight was measured during the experimental period in the sham group (diamonds), OVX group (squares) and OVX rats with oral administration of HEP extract at 110 mg kg−1 d−1 (open triangles) or 17-β estradiol at 2 mg kg−1 d−1 (filled triangles). (B) Uterine weight was measured in the baseline group and 4 months after operation in the sham group, OVX group, OVX rats treated with 110 mg kg−1 d−1 of HEP extract and OVX rats treated with 2 mg kg−1 d−1 of 17-β estradiol. Data are expressed as mean ± SEM. *P < 0.05, **P < 0.01, versus sham control; ##P < 0.01, versus OVX group.
<b>Figure 3</b>
Figure 3
Bone histomorphometric analysis of rat tibia. The images of the metaphyseal trabeculae of the sham (A), OVX (B), E2 (C) and HEP (D) groups.
<b>Figure 4</b>
Figure 4
Effects of HEP extracts on cell proliferation and ALP activity in rat osteoblast-like UMR 106 cells. (A) Cell proliferation of UMR 106 cells upon HEP extract treatment. UMR 106 cells were seeded in a 96-well plate at a density of 2500 cells per well. After starvation for 24 h, different concentrations (50, 100 and 200 μg ml−1) of HEP extract were added, and treated for 24 or 48 h. Vehicle-treated cells served as control (ctrl). Cell number was determined by MTT assay. Results were obtained from two independent experiments and expressed as mean ± SEM. **P < 0.01, versus control for n = 6. (B) ALP activity of UMR 106 cells upon HEP extract treatment. UMR 106 cells were seeded in 24-well plates at a density of 100 000 cells per well. After starvation, 50, 100 and 200 μg ml−1 of HEP were added, and treated for 24 h before ALP activity determination. Vehicle-treated cells served as control (ctrl). The ALP activity was corrected for the amount of protein used and expressed as U l−1 μg−1. Results were obtained from two independent experiments and expressed as mean ± SEM. **P < 0.01 versus control for n = 3.
<b>Figure 5</b>
Figure 5
Effect of HEP on OPG and receptor activator of NF-κB ligand (RANKL) mRNA expression in rat osteoblast-like UMR 106 cells. UMR 106 cells were treated with 100 μg ml−1 and 200 μg ml−1 HEP extract or its vehicle (ctrl) for 48 h. Total RNA was isolated and subjected to semi-quantitative RT–PCR analysis of OPG (A) and RANKL (B) mRNA expression under the conditions described in ‘Methods’. (C) The ratio of OPG/RANKL was normalized by an internal control gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Results were obtained from two independent experiments and expressed as mean ± SEM. *P < 0.05, **P < 0.01, versus control for n = 3.
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