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

다국어 초록 (Multilingual Abstract)

2,3,5,4’-Tetrahydroxystilbene-2-O-β-D-glucoside (TSG), an active polyphenolic component of Polygonum multiflorum, exhibits many pharmacological activities including antioxidative, anti-inflammatory, 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 bone 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 as compared with the OVX mice. Additionally, the serum bone alkaline phosphatase (BALP) levels in the TSGtreated OVX mice were significantly increased compared with the OVX mice without treatment of TSG, while the tartrate-resistant acid phosphatase (TRAP)
activity was significantly reduced. Furthermore, the OVX mice treated with TSG showed a significantly reduced bone loss on micro-computed tomography (CT) analysis compared to the untreated OVX mice. Consequently, bone destruction in osteoporotic mice as a result of ovariectomy was inhibited by 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.

목차 (Table of Contents)

• Ⅰ. Introduction 1
• Ⅱ. Materials and Methods 4
• 1. Reagents. 4
• 2. Cell cultures and osteoclast differentiation. 4
• 3. Cell Cytotoxicity Assay 5
• 4. Tartrate-Resistant acid Phosphatase (TRAP) Staining Assay 5
• 5. Osteoporosis Mice Model and Experimental Treatments. 5
• 6. Analyses of Serum Ca, IP, ALP, and TCHO. 6
• 7. Analyses of Serum Tartrate-Resistant Acid Phosphatase (TRAP) and BALP. 7
• 8. Measurement of Bone Structure Using Micro-Computed Tomography (CT). 7
• 9. Bone and Uterus Histological Analysis. 8
• 10. Statistical analysis 9
• Ⅲ. Results 10
• 1. Effects of TSG on osteoclast differentiation and cytotoxicity from BMMs 10
• 2. Effects of TSG on Body Weight of Ovariectomized (OVX) Mice 10
• 3. Effect of TSG on Organ (Uterus, Spleen, and Thymus) Weight in OVX Mice 11
• 4. Effect of TSG on Bone Weight and Length in OVX Mice. 11
• 5. Effect of TSG on Serum Biochemical Markers (Calcium (Ca), Inorganic Phosphorus (IP), Alkaline Phosphatase (ALP), and Total Cholesterol (TCHO)) in OVX Mice. 12
• 6. Effect of TSG on Serum Tartrate-Resistant Acid Phosphatase (TRAP) and Bone Specific Alkaline Phosphatase (BALP) in OVX Mice 13
• 7. Effect of TSG on Bone Structure Using Micro-Computed Tomography (Micro-CT) in OVX Mice 13
• 8. Effect of TSG on Histology of Bones and Uteri of OVX Mice. 15
• Ⅳ. Discussion. 29
• Ⅴ. Conclusion. 34
• Ⅵ. References. 38