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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (12): 1419-1426.doi: 10.3736/jcim20121214

• Original Experimental Research • Previous Articles     Next Articles

Antiosteoporotic effects of benzylbenzoate glucosides from Curculigo orchioides in ovariectomized rats

Lei Liu1,2, Yuan-hui Guo1,3, Hai-liang Xin4, Yan Nie1,2, Ting Han1, Lu-ping Qin1(), Qiao-yan Zhang1()   

  1. 1. Department of Pharmacogonosy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
    2. Department of Pharmacogonosy, Fujian University of Traditional Chinese Medicine, Fuzhou 350108, Fujian Province, China
    3. Department of Pharmacy, No. 91892 Hospital of PLA, Sansha 573100, Hainan Province, China
    4. School of Traditional Chinese Medicine, Second Military Medical University, Shanghai 200433, China
  • Received:2012-05-19 Accepted:2012-06-20 Online:2012-12-20 Published:2018-12-15

Objective: To evaluate the antiosteoporotic effects of benzylbenzoate glucosides from Curculigo orchioides (COBG) in ovariectomized (OVX) rats.
Methods: A total of 70 female Sprague-Dawley rats were assigned to sham-operated and OVX model groups. The OVX rats were further divided into six subgroups treated by gavage with vehicle, 1 mg/kg of nylestriol, 6, 18 and 54 mg/kg of COBG and 3.0 g/kg of ethanol extract of Curculigo orchioides respectively for 12 weeks. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry. The sections of tibia were prepared for histomorphometric analysis. The biomarkers in serum and urine were determined using reagent kits.
Results: Ovariectomy induced the bone loss and microarchitectural deterioration of bone tissue with the activities of increased serum alkaline phosphatase and loss of calcium through the excretion in urine, and decreased levels of antioxidant in serum (P<0.05, P<0.01). Administration of 6, 18 and 54 mg/kg of COBG significantly increased the BMD, improved the microarchitecture of bone tissue, prevented the depletion of antioxidant enzymes like superoxide dismutase and glutathione peroxidase, inhibited the increase of malondialdehyde in serum and reduced the excretion of urine calcium in OVX rats (P<0.05, P<0.01).
Conclusion: COBG could prevent the bone loss through improving the antioxidant status, which offers a potential new therapeutic drug for postmenopausal osteoporosis.

Key words: Curculigo orchioides, ovariectomy, osteoporosis,postmenopausal, antioxidant, rats,Sprague-Dawley

Figure 1

The chemical structure of two benzylbenzoate glucosidesThe present study aimed to evaluate the effects of COBG on bone metabolism and oxidative status in ovariectomized (OVX) rats by measuring the bone density and the change in bone histological morphology, serum biochemical parameters including levels of calcium and phosphorus, serum alkaline phosphatase (ALP) activity, malondialdehyde (MDA) content and the depletion of some enzymatic antioxidants such as SOD, GSH-Px, and nitric oxide synthase (NOS)."


Group n Initial body weight Final body weight Uterine weight
Sham-operated 10 241.8±6.2 287.3±10.0 1.17±0.24
OVX model 10 252.4±18.7 346.4±24.4** 0.26±0.06**
OVX plus nylestriol 10 256.3±21.6 289.1±12.1△△ 0.74±0.24△△
OVX plus 3 g/kg of COE 10 256.9±14.4 344.1±16.2** 0.27±0.08**
OVX plus 6 mg/kg of COBG 10 258.5±24.7 361.1±30.0** 0.30±0.05**
OVX plus 18 mg/kg of COBG 10 251.3±21.9 326.3±41.7** 0.27±0.07**
OVX plus 54 mg/kg of COBG 10 254.8±12.8 329.3±36.0** 0.32±0.09**


Group n Serum calcium
Serum phosphorus
Urine calcium/
Cr (mmol/mmol)
Urine phosphorus/
Cr (mmol/mmol)
Serum ALP
Sham-operated 10 2.80±0.25 3.06±0.61 0.40±0.13 3.92±1.29 33.9±5.3
OVX model 10 2.61±0.15 2.52±0.59 1.07±0.34** 4.80±1.20 46.5±10.6*
OVX plus nylestriol 10 2.70±0.07 2.61±0.40 0.70±0.29 5.73±1.66 33.9±3.4
OVX plus 3 g/kg of COE 10 2.60±0.06 2.56±0.34 0.47±0.12△△ 5.46±1.91 52.0±14.0
OVX plus 6 mg/kg of COBG 10 2.78±0.13 2.66±0.35 0.40±0.10△△ 3.58±1.13 51.3±3.0
OVX plus 18 mg/kg of COBG 10 2.70±0.14 2.79±0.34 0.63±0.28 4.18±1.18 43.4±5.2
OVX plus 54 mg/kg of COBG 10 2.67±0.08 2.44±0.35 0.53±0.20△△ 4.27±1.3 48.0±11.1

Figure 2

Effects of COBG on bone mineral density in OVX rats. Data are expressed as mean±standard deviation, n=10; **P<0.01, vs sham-operated group; △△P<0.01, vs OVX model group. t-BMD: tibia bone mineral density; OVX: ovariectomized; COE: Curculigo orchioides extracts; COBG: benzylbenzoate glucosides from Curculigo orchioides"

Figure 3

Histopathology of the cancellous bone of tibia stained with Goldner’s Trichrome (Light microscopy, ×40) A: Normal compactness of the diaphysis and competent trabeculae in the sham-operated group; B: Sparse, uniform thinning of the trabeculae resulting in widening of intertrabecular spaces in the OVX model animals; C: Nylestriol treatment group; D: 6 mg/kg of COBG group; E: 18 mg/kg of COBG group exhibited significant restorative progress with mineralization along with quite well distributed osteocytes, where uniform trabeculae with dense matrix and shaft size were observed; F: 54 mg/kg of COBG group showed nearly complete recovery with essential features of normal bone."


Group n Percent trabecular
area (%)
thickness (μm)
number (/mm)
separation (μm)
Sham-operated 10 20.4±3.7 61.5±6.1 3.69±0.51 214.93±47.2
OVX model 10 7.6±3.0** 61.9±4.6 1.22±0.51** 1026.7±584.5**
OVX plus nylestriol 10 18.0±5.3△△ 59.3±7.2 2.85±0.95△△ 327.9±128.1△△
OVX plus 6 mg/kg of COBG 10 10.9±7.1 60.0±7.6 1.89±1.37 772.6±512.6
OVX plus 18 mg/kg of COBG 10 12.3±7.0 63.0±6.6 1.64±1.29 535.6±309.9
OVX plus 54 mg/kg of COBG 10 14.3±5.5△△ 60.8±7.0 2.76±1.08△△ 541.8±281.6


Group n Label
perimeters (%)
rate (μm/d)
(percent per year )
Osteoclast number
Sham-operated 10 13.8±6.0 0.63±0.30 93.7±53.5 7.47±3.25
OVX model 10 31.9±21.5* 1.21±0.32** 360.5±47.7** 18.84±3.67**
OVX plus nylestriol 10 15.5±7.5 0.74±0.20△△ 109.5±35.9△△ 8.84±4.24△△
OVX plus 6 mg/kg of COBG 10 30.0±20.9 1.06±0.20 327.0±58.5 14.74±6.53
OVX plus 18 mg/kg of COBG 10 34.6±10.0 0.94±0.09 334.3±32.6 11.28±2.80△△
OVX plus 54 mg/kg of COBG 10 32.5± 5.8 1.18±0.19 345.5±46.6 9.75±2.63△△

Figure 4

Effects of COBG on serum SOD, GSH-Px, NOS and MDA in OVX rats Data are expressed as mean±standard deviation, n=10; *P<0.05, **P<0.01, vs sham-operated group; △P<0.05, △△P<0.01, vs OVX model group. A: Effects of COBG on serum SOD in each group; B: Effects of COBG on serum GSH-Px in each group; C: Effects of COBG on serum NOS in OVX rats; D: Effects of COBG on serum MDA in each group. SOD: super oxygen dehydrogenise; NOS: nitric oxide synthase; MDA: malondialdehyde; GSH-Px: glutathione peroxidase; OVX: ovariectomized; COBG: benzylbenzoate glucosides from Curculigo orchioides; COE: curculigo orchioides extracts."

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