Anat Cell Biol.  2017 Sep;50(3):219-229. 10.5115/acb.2017.50.3.219.

Protective effect of Rhus verniciflua Stokes extract in an experimental model of post-menopausal osteoporosis

• Jeong JH
• An JH
• Yang H
• Kim DK
• Lee NS
• Jeong YG
• Na CS
• Na DS
• Dong MS
• Han SY

• KMID: 2390484
• DOI: http://doi.org/10.5115/acb.2017.50.3.219

Abstract

Post-menopausal osteoporosis (PMO) is a major global human health concern. Owing to the need for therapeutic drugs without side effects, natural extracts containing various polyphenolic compounds that may exert estrogenic effects have been studied in depth. Rhus verniciflua Stokes (RVS), which has been used as a traditional herbal medicine for centuries in Korea, was recently revealed to exert estrogenic effects attributable to its bioactive ingredients sulfuretin and butein, which have strong estrogen receptor-binding affinities. In this study, the protective potential of RVS in PMO was evaluated by using an experimental animal model of PMO, which was established by ovariectomy (OVX) of female Sprague Dawley rats. The oral administration of RVS at 20 mg/kg or 100 mg/kg for 8 weeks markedly protected against OVX-induced atrophy of the uterine tube and reversed the elevation in the ratio of serum receptor activator of nuclear factor-κB ligand to osteoprotegerin, which is a marker of disease severity. In addition, RVS inhibited OVX-induced tibia bone loss, activated osteogenic activity, and suppressed osteoclastic activity in the tibial epiphyseal plate, a region of bone remodeling. Collectively, these factors indicated that the oral intake of RVS might be beneficial for the prevention of PMO.

Keyword

Rhus verniciflua Stokes; Postmenopausal osteoporosis; Ovariectomy; Bone remodeling

MeSH Terms

Administration, Oral
Atrophy
Bone Remodeling
Estrogens
Fallopian Tubes
Female
Growth Plate
Herbal Medicine
Humans
Korea
Models, Animal
Models, Theoretical*
Osteoclasts
Osteoporosis, Postmenopausal*
Osteoprotegerin
Ovariectomy
Rats, Sprague-Dawley
Rhus*
Tibia
Estrogens
Osteoprotegerin

Figure

• 

  Fig. 1 Protective effect of Rhus verniciflua Stokes (RVS) on ovariectomy-induced atrophy of the uterine tubes. Rats (n=12 in each group) were intragastrically treated with or without RVS (20 mg/kg for RVS at a low dose [RVS-L] and 100 mg/kg for RVS at a high dose [RVS-H] group) for 8 weeks after ovariectomy. The sham-surgery (Sham) group was subjected to laparotomy, but not ovariectomy. The ovariectomy (OVX) group was administered distilled water as a vehicle. The photomicrographs are representative gross morphologies (A, upper panel) and H&E-stained transverse sections (A, lower panel) of the excised uterine tubes of each group. The average wet weight (B) and thickness (C) of the uterine tubes of each group were quantitatively assessed. The data in all graphs are expressed as the mean±SEM (*P<0.05 and ***P<0.001 vs. Sham, #P<0.05, ##P<0.01, and ###P<0.001 vs. OVX).

• 

  Fig. 2 Protective effects of Rhus verniciflua Stokes (RVS) on ovariectomy-induced deterioration of rat tibial microstructure. Rats (n=12 in each group) were intragastrically treated with or without RVS (20 mg/kg for RVS at a low dose [RVS-L] and 100 mg/kg for RVS at a high dose [RVS-H]) for 8 weeks after ovariectomy. The sham-surgery (Sham) group was subjected to laparotomy, but not ovariectomy. The ovariectomy (OVX) group was administered distilled water as a vehicle. After treatment of RVS for 8 weeks, both tibiae of each rat were excised. After the decalcification step, the bone microstructure of proximal tibia was analyzed by microcomputed tomography (µCT). Representative µCT images are shown in panel (A) and the parameters such as bone volume per tissue volume (BV/TV) (B), trabecular thickness (Tb.Th) (C), trabecular number (Tb.N) (D), trabecular separation (Tb.Sp) (E), trabecular pattern factor (Tb.Pf) (F), and structure model index (SMI) (G) were analyzed. The data in all graphs are expressed as the mean±SEM (*P<0.05, **P<0.01, and ***P<0.001 vs. Sham; #P<0.05, ##P<0.01, and ###P<0.001 vs. OVX).

• 

  Fig. 3 Suppressive effect of Rhus verniciflua Stokes (RVS) on ovariectomy-induced elevation of the serum receptor activator of nuclear factor-κB ligand (RANKL)/osteoprotegerin (OPG) ratio. Rats (n=12 in each group) were intragastrically treated with or without RVS (20 mg/kg for RVS at a low dose [RVS-L] and 100 mg/kg for RVS at a high dose [RVS-H] group) for 8 weeks after ovariectomy. The sham-surgery (Sham) group was subjected to laparotomy, but not ovariectomy. The ovariectomy (OVX) group was administered distilled water as a vehicle. After RVS treatment for 8 weeks, the serum levels of OPG (A) and RANKL (B) were determined by using enzyme-linked immunosorbent assay kits and the ratio of RANKL to OPG (C) was calculated. The data in all graphs are presented as the mean±SEM (*P<0.05 and **P<0.01 vs. Sham, #P<0.05 vs. OVX).

• 

  Fig. 4 Preserved or enhanced activity of bone remodeling in tibial epiphyseal plate by Rhus verniciflua Stokes (RVS) treatment in ovariectomy-induced osteoporotic rats. Rats (n=12 in each group) were intragastrically treated with or without RVS (20 mg/kg for RVS at a low dose [RVS-L] and 100 mg/kg for RVS at a high dose [RVS-H] group) for 8 weeks after ovariectomy. The sham-surgery (Sham) group was subjected to laparotomy, but not ovariectomy. The ovariectomy (OVX) group was administered distilled water as a vehicle. After RVS treatment for 8 weeks, both tibiae of each rat were excised. After a decalcification step, the histological morphology in the epiphyseal plate of tibia was observed after being stained with H&E (A, ×100, a representative image) or hematoxylin (B, ×400, the magnified regions of corresponding yellow rectangular boxes in A). Yellow arrowheads indicate the discontinued trabecular meshwork. The average length of resting zone (RZ) and proliferation zone (PZ) in the tibial epiphyseal plate of each group is quantitatively expressed in panel (C). The data in all graphs are expressed as the mean±SEM (*P<0.05 and **P<0.01 vs. Sham, #P<0.05 and ##P<0.01 vs. OVX).

• 

  Fig. 5 Inhibition of osteoclastic activity in the tibial epiphyseal plate by Rhus verniciflua Stokes (RVS) treatment in ovariectomy-induced osteoporotic rats. Rats (n=12 in each group) were intragastrically treated with or without RVS (20 mg/kg for RVS at a low dose [RVS-L] and 100 mg/kg for RVS at a high dose [RVS-H] group) for 8 weeks after ovariectomy. The sham-surgery (Sham) group was subjected to laparotomy, but not ovariectomy. The ovariectomy (OVX) group was administered distilled water as a vehicle. After treatment of RVS for 8 weeks, both tibiae of each rat were excised. As shown in the representative images (A, ×100) and the quantitative graph (B), after the decalcification step, the activity of osteoclasts in tibia epiphyseal plate of each group was quantitatively analyzed by the analysis of the optical density of the brown-colored immunoreaction (indicated by white arrowheads) after immunohistochemical staining with tartrate-resistant acid phosphatase (TRAP). The data are expressed as the mean±SEM (*P<0.05 and ***P<0.001 vs. Sham, ##P<0.01 vs. OVX).

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