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. 2018 Aug 28;19(9):2554.
doi: 10.3390/ijms19092554.

Protective Effects of 2,3,5,4'-Tetrahydroxystilbene-2- O-β-d-glucoside on Ovariectomy Induced Osteoporosis Mouse Model

Su-Jin Kim  1 , Yun-Ho Hwang  2 , Seul-Ki Mun  3 , Seong-Gyeol Hong  4 , Kwang-Jin Kim  5 , Kyung-Yun Kang  6 , Young-Jin Son  7 , Sung-Tae Yee  8
Affiliations

Protective Effects of 2,3,5,4'-Tetrahydroxystilbene-2- O-β-d-glucoside on Ovariectomy Induced Osteoporosis Mouse Model

Su-Jin Kim et al. Int J Mol Sci. .

Abstract

2,3,5,4'-Tetrahydroxystilbene-2-O-β-d-glucoside (TSG), an active polyphenolic component of Polygonum multiflorum, exhibits many pharmacological activities including antioxidant, anti-inflammation, and anti-aging effects. A previous study demonstrated that TSG protected MC3T3-E1 cells from hydrogen peroxide (H₂O₂) induced cell damage and the inhibition of osteoblastic differentiation. However, no studies have investigated the prevention of ovariectomy-induced bone loss in mice. Therefore, we investigated the effects of TSG on bone loss in ovariectomized mice (OVX). Treatment with TSG (1 and 3 μg/g; i.p.) for six weeks positively affected body weight, uterine weight, organ weight, bone length, and weight change because of estrogen deficiency. The levels of the serum biochemical markers of calcium (Ca), inorganic phosphorus (IP), alkaline phosphatase (ALP), and total cholesterol (TCHO) decreased in the TSG-treated mice when compared with the OVX mice. Additionally, the serum bone alkaline phosphatase (BALP) levels in the TSG-treated OVX mice were significantly increased compared with the OVX mice, while the tartrate-resistant acid phosphatase (TRAP) activity was significantly reduced. Furthermore, the OVX mice treated with TSG showed a significantly reduced bone loss compared to the untreated OVX mice upon micro-computed tomography (CT) analysis. Consequently, bone destruction in osteoporotic mice as a result of ovariectomy was inhibited by the administration of TSG. These findings indicate that TSG effectively prevents bone loss in OVX mice; therefore, it can be considered as a potential therapeutic for the treatment of postmenopausal osteoporosis.

Keywords: 2,3,5,4′-tetrahydroxystilbene-2-O-β-d-glucoside (TSG); bone loss; menopause; osteoporosis; ovariectomy.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effect of TSG on body weight. Body weights were measured at weekly intervals and the effect of TSG was seen at six weeks. a, b, and c: The means not sharing a common letter are significantly different among group at p < 0.05 by one-way analysis of variance (ANOVA) with Duncan’s multiple-range test. SHAM (sham-operated control group); OVX (ovariectomized group); TSG1 (1 μg OVX TSG group); TSG 3 (3 μg OVX TSG group).
Figure 2
Figure 2
Effect of TSG on uterus weight. The C3H/HeN mice were treated with TSG for 6 weeks, and the uteri were harvested 24 h after the last treatment. (A) The uterus was photographed with a digital camera and (B) weighed. a, b, and c: The means not sharing a common letter are significantly different among group at p < 0.05 by one-way ANOVA with Duncan’s multiple-range test.
Figure 3
Figure 3
Effect of TSG on the serum biochemical markers. In the control, the SHAM-operated mice and OVX mice with or without the administration of TSG (1 and 3 µg/g/day, I.P) for six weeks, the serum (A) calcium, (B) phosphorus, (C) alkaline phosphatase, and (D) total cholesterol were determined using a diagnostic slide. a, b, and c: The means not sharing a common letter are significantly different among the groups at p < 0.05 by one-way ANOVA with Duncan’s multiple-range test.
Figure 4
Figure 4
Effect of TSG on (A) tartrate-resistant acid phosphatase (TRAP) and (B) bone specific alkaline phosphatase (BALP) in the serum after six weeks of treatment. a, b, and c: The means not sharing a common letter are significantly different among groups at p < 0.05 by one-way ANOVA with Duncan’s multiple-range test.
Figure 5
Figure 5
Effect of TSG on the trabecular morphometric parameters in the distal femur of C3H/HeN mice. The mice were treated with vehicle and TSG (1 and 3 µg/g/day, physiological phosphate [IP]) for six weeks. (A) The representative two-dimensional (2D) images and three dimensional (3D) images of the femur epiphysis, (B) tissue volume, (C) bone volume, (D) bone volume/tissue volume, (E) bone surface, (F) bone surface/tissue volume, (G) trabecular pattern factor, (H) structure model index, (I) trabecular thickness, (J) trabecular number, and (K) trabecular separation, as analyzed by the micro-computed tomography (CT) Skyscan CTAn software. a, b, and c: The means not sharing a common letter are significantly different among groups at p < 0.05 by one-way ANOVA with Duncan’s multiple-range test.
Figure 5
Figure 5
Effect of TSG on the trabecular morphometric parameters in the distal femur of C3H/HeN mice. The mice were treated with vehicle and TSG (1 and 3 µg/g/day, physiological phosphate [IP]) for six weeks. (A) The representative two-dimensional (2D) images and three dimensional (3D) images of the femur epiphysis, (B) tissue volume, (C) bone volume, (D) bone volume/tissue volume, (E) bone surface, (F) bone surface/tissue volume, (G) trabecular pattern factor, (H) structure model index, (I) trabecular thickness, (J) trabecular number, and (K) trabecular separation, as analyzed by the micro-computed tomography (CT) Skyscan CTAn software. a, b, and c: The means not sharing a common letter are significantly different among groups at p < 0.05 by one-way ANOVA with Duncan’s multiple-range test.
Figure 6
Figure 6
Effect of TSG on trabecular morphometric parameters in the proximal tibia of the C3H/HeN mice. The mice were treated with vehicle and TSG (1 and 3 µg/g/day, IP) for six weeks. (A) Representative two-dimensional (2D) images and three dimensional (3D) images of the tibia epiphysis, (B) tissue volume (TV), (C) bone volume (BV), (D) bone volume/tissue volume, (E) bone surface, (F) bone surface/tissue volume, (G) trabecular pattern factor, (H) structure model index, (I) trabecular thickness, (J) trabecular number, and (K) trabecular separation as analyzed by the micro-CT Skyscan CTAn software. a, b, c, and d: The means not sharing a common letter are significantly different among groups at p < 0.05 by one-way ANOVA with Duncan’s multiple-range test.
Figure 7
Figure 7
Effect of TSG on the bone tissue of the trabecular in the distal femur of C3H/HeN mice. The mice were treated with vehicle and TSG (1 and 3 µg/g/day, IP) for six weeks. (A) Histological analysis of distal femur with hematoxylin and eosin (H and E) and tartrate-resistant acid phosphatase (TRAP), Masson’s trichrome staining (400× magnification); (B) trabecular bone area; (C) TRAP positive cells and (D) collagen in the femur were analyzed using the Image J program. a, b, c, d, and e: The means not sharing a common letter are significantly different among groups at p < 0.05 by one-way ANOVA with Duncan’s multiple-range test.
Figure 8
Figure 8
Effect of TSG on the trabecular bone tissue in the proximal tibia of C3H/HeN mice. Mice were treated with vehicle and TSG (1 and 3 µg/g/day, I.P.) for six weeks. (A) Histological analysis of the proximal tibia with hematoxylin and eosin (H and E) and tartrate-resistant acid phosphatase (TRAP), Masson’s trichrome staining (400× magnification); (B) trabecular bone area; (C) TRAP positive cell and (D) collagen in tibia were analyzed by the Image J program. a, b, c, d, and e: The means not sharing a common letter are significantly different among groups at p < 0.05 by one-way ANOVA with Duncan’s multiple-range test.
Figure 9
Figure 9
Effect of TSG on uterus tissue in C3H/HeN mice. Mice were treated with vehicle and TSG (1 and 3 μg/g/day, I.P.) for six weeks. Histological changes in the uterus were performed via H and E staining.
Figure 10
Figure 10
(A) The molecular structure of 2,3,5,4’-Tetrahydroxystilbene-2-O-β-d-glucoside (TSG); (B) experimental protocol for the induction and treatment of osteoporosis, along with the treatment scheme. OVX—ovariectomized mice; E2—estradiol.
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