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. 2016 Sep 10:5:262-273.
doi: 10.1016/j.bonr.2016.09.001. eCollection 2016 Dec.

Effects of total flavonoids from Drynariae Rhizoma prevent bone loss in vivo and in vitro

Shuang-Hong Song  1   2   3 , Yuan-Kun Zhai  4 , Cui-Qin Li  2   3 , Qian Yu  2   3 , Yi Lu  1 , Yuan Zhang  2   3 , Wen-Ping Hua  2   3 , Zhe-Zhi Wang  2   3 , Peng Shang  1
Affiliations

Effects of total flavonoids from Drynariae Rhizoma prevent bone loss in vivo and in vitro

"VSports最新版本" Shuang-Hong Song et al. Bone Rep. .

Abstract

Estrogen deficiency is one of the major causes of osteoporosis in postmenopausal women. Drynariae Rhizoma is a widely used traditional Chinese medicine for the treatment of bone diseases. In this study, we investigated the therapeutic effects of the total Drynariae Rhizoma flavonoids (DRTF) on estrogen deficiency-induced bone loss using an ovariectomized rat model and osteoblast-like MC3T3-E1 cells. Our results indicated that DRTF produced osteo-protective effects on the ovariectomized rats in terms of bone loss reduction, including decreased levels of bone turnover markers, enhanced biomechanical femur strength and trabecular bone microarchitecture deterioration prevention. In vitro experiments revealed that the actions of DRTF on regulating osteoblastic activities were mediated by the estrogen receptor (ER) dependent pathway. Our data also demonstrated that DRTF inhibited osteoclastogenesis via up-regulating osteoprotegrin (OPG), as well as down-regulating receptor activator of NF-κB ligand (RANKL) expression. In conclusion, this study indicated that DRTF treatment effectively suppressed bone mass loss in an ovariectomized rat model, and in vitro evidence suggested that the effects were exerted through actions on both osteoblasts and osteoclasts VSports手机版. .

Keywords: Drynariae Rhizoma; Osteoblast; Osteoclast; Osteoporosis; Ovariectomy. V体育安卓版.

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Figures (V体育官网)

Fig. 1
Fig. 1
The structures of naringin (A) and neoeriocitrin (B); the HPLC chromatograms of the reference substances (C) and DRTF (D). Peak 1 is neoeriocitrion, peak 2 is naringin; the primary mass spectrograms of naringin (E) and neoeriocitrion (F).
Fig. 2
Fig. 2
Effects of DRTF or E2 on body weights and organ indices in OVX rats. (A) The body weights of the animals were recorded every 2 weeks during the experimental period. Values are mean ± SD (n = 8), *p < 0.05, **p < 0.01compared with sham; #p < 0.05, ##p < 0.01 compared with OVX. (B) Uteri were isolated and weighed after euthanization, the uterus index was determined as the uterus weight divided by body weight. Values are mean ± SD (n = 8), *p < 0.001, ***p < 0.001 compared with sham; ^^^p < 0.001 compared with E2 treatment. (C) HE dyed sections of rat uteri obtained from (a) Sham, (b) OVX, (c) E2, (d) DRTF25, (e) DRTF75, and (f) DRTF225 treated animals. (D) The heart, liver, spleen, lung, kidney, brain and thymus were isolated and weighed after the animals were sacrificed and the organ index was represented as organ weight divided by body weight.
Fig. 3
Fig. 3
Effects of 12-week treatment on the BMD and BMC in the femur of OVX rats by DEXA. Values are mean ± SD (n = 8), ***p < 0.001 different from Sham; #p < 0.05, ##p < 0.01 different from OVX.
Fig. 4
Fig. 4
Representative sample from each group: 3D architecture of trabecular bone within the distal metaphyseal femur region: (A) Sham, (B) OVX, (C) E2, (D) DRTF25, (E) DRTF75, (F) DRTF225.
Fig. 5
Fig. 5
Effects of DRTF on the proliferation and differentiation of MC3T3-E1 cells: (A) Cell proliferation of MC3T3-E1 was evaluated by the MTT method. Values are mean ± SD (n = 8), **p < 0.01 vs. control. (B) ALP activity was assessed using Alkaline Phosphate Assay Kit. Values are mean ± SD (n = 8), *p < 0.05, **p < 0.01 vs. control.
Fig. 6
Fig. 6
Mineralized nodules formation of MC3T3-E1 cells cultured with osteogenic medium. (A) Representative images of mineralized nodules stained by Alizarin Red S, (a) DMEM control, (b) DRTF (12.5 μg/ml) treatment, bar: 100 μm. (B) Quantitative analysis of the area of mineralized nodules using Image J software. Values are mean ± SD (n = 3), **p < 0.01 vs. control.
Fig. 7
Fig. 7
Effects of DRTF on OPG and RANKL mRNA expression in MC3T3-E1 cells. MC3T3-E1 cells were treated with vehicle (control), E2 and different concentrations of DRTF (2.5, 12.5 and 62.5 μg/ml) for 12 h. Total RNA was isolated and real-time RT-PCR was performed to determine the mRNA expression levels of OPG (A) and RANKL (B), which were normalized to that of GAPDH. The results are expressed as the mean ± SD (n = 3). (C) Effect of DRTF on the OPG/RANKL message level ratio in MC3T3-E1 cells. Data are presented as the mean ratio of OPG:RANKL expression ± SD (n = 3). *p < 0.05, **p < 0.01 vs. control.
Fig. 8
Fig. 8
Effects of DRTF on the OPG and RANKL protein levels in MC3T3-E1 cells. (A) Western blot analysis of OPG and RANKL expression after treatment of MC3T3-E1 cells with E2 or DRTF (12.5 μm/ml) for 48 h. (B) “Quantity one” from Bio-Rad was used to analyze the western blot results, the values represent mean ± SD (n = 3), *p < 0.05, **p < 0.01 vs. control.
Fig. 9
Fig. 9
Effects of DRTF on ERα and ERβ mRNA expression in MC3T3-E1 cells. MC3T3-E1 cells were treated with vehicle (control), E2 and different concentrations of DRTF (2.5, 12.5 and 62.5 μg/ml) for 12 h. Total RNA was isolated and real-time RT-PCR was performed to determine the mRNA expressions levers of ERα (A) and ERβ (B), which were normalized to that of GAPDH. The results were expressed as the mean ± SD (n = 3), *p < 0.05, **p < 0.01 vs. control.
Fig. 10
Fig. 10
Effects of DRTF on the ERα and ERβ protein level in MC3T3-E1 cells. (A) Western blot analysis of ERα and ERβ expressions after treatment of MC3T3-E1 cells with E2 or DRTF (62.5 μm/ml) for 48 h. (B) “Quantity one” from Bio-Rad was used to analyze the western blotting results. The values were expressed as mean ± SD (n = 3), *p < 0.05, **p < 0.01 vs. control.
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References

    1. Al-Anazi A.F., Qureshi V.F., Javaid K., Qureshi S. Preventive effects of phytoestrogens against postmenopausal osteoporosis as compared to the available therapeutic choices: an overview. J. Nat. Sci. Biol. Med. 2011;2:154–163. - PMC - PubMed
    1. Banu J., Varela E., Fernandes G. Alternative therapies for the prevention and treatment of osteoporosis. Nutr. Rev. 2012;70:22–40. - V体育官网 - PubMed
    1. Bonjour J.P., Benoit V., Rousseau B., Souberbielle J.C. Consumption of vitamin D-and calcium-fortified soft white cheese lowers the biochemical marker of bone resorption TRAP 5b in postmenopausal women at moderate risk of osteoporosis fracture. J. Nutr. 2012;142:698–703. - PubMed
    1. Boonchird C., Mahapanichkul T., Cherdshewasart W. Differential binding with ERalpha and ERbeta of the phytoestrogen-rich plant Pueraria mirifica. Braz. J. Med. Biol. Res. 2010;43:195–200. - PubMed
    1. Bord S., Ireland D.C., Beavan S.R., Compston J.E. The effects of estrogen on osteoprotegerin, RANKL, and estrogen receptor expression in human osteoblasts. Bone. 2003;32:136–141. - PubMed

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