Nutr Res Pract.  2015 Oct;9(5):459-465. 10.4162/nrp.2015.9.5.459.

St. John's Wort (Hypericum perforatum) stimulates human osteoblastic MG-63 cell proliferation and attenuates trabecular bone loss induced by ovariectomy

• You MK
• Kim DW
• Jeong KS
• Bang MA
• Kim HS
• Rhuy J
• Kim HA

• KMID: 2313865
• DOI: http://doi.org/10.4162/nrp.2015.9.5.459

Abstract

BACKGROUND
/OBJECFTIVES: The effect of St. John's Wort extract (SJW) on MG-63 cell proliferation and trabecular bone loss induced by ovariectomy was examined.
MATERIALS/METHODS
Proliferation, expression of estrogen receptor (ER) alpha and ER beta, and gene expressions of osteoprotegerin (OPG), osteocalcin (OC) and alkaline phosphatase (ALP) were examined in MG-63 cells treated with or without SJW. Ovariectomized rats were treated with SJW at the dose of 100 or 200 mg/kg/day, beta-estradiol-3-benzoate (E2), or vehicle only (OVX-C), and sham operated rats were treated with vehicle only (Sham-C). Serum ALP and C-telopeptide (CTX), and femoral trabecular bone loss were examined.
RESULTS
SJW increased MG-63 cell proliferation and expression of ER alpha and ER beta, and positive effect was shown on gene expressions of ALP, OC and OPG. SJW also showed estrogen like effect on bone associated with slowing down in trabecular bone loss. Histopathology by H&E showed rats treated with SJW displayed denser structure in metaphyseal region of distal femur compared with rats in OVX-C. SJW was shown to reduce serum CTX in OVX rats.
CONCLUSION
The present study provides new insight in preventing estrogen deficiency induced bone loss of SJW and possibility for its application in bone health supplement.

Keyword

St. John's Wort; MG-63 cells; proliferation; ovariectomy; osteoporosis

MeSH Terms

Alkaline Phosphatase
Animals
Cell Proliferation*
Estrogens
Female
Femur
Gene Expression
Humans*
Hypericum*
Osteoblasts*
Osteocalcin
Osteoporosis
Osteoprotegerin
Ovariectomy*
Rats
Alkaline Phosphatase
Estrogens
Osteocalcin
Osteoprotegerin

Figure

• 

  Fig. 1 Effect of St. John's Wort extract on cytotoxicity in MG 63 cells. Values are mean ± SD. Stand error bars represent three independent experiments and each experiment was triplicated. Means with the same letter are not significantly different by Duncan's multiple range test (P < 0.05). SJW: 70% ethanol extract of St. John's Wort

• 

  Fig. 2 Effect of St. John's Wort extract on proliferation in MG 63 cells. Values are mean ± SD. Stand error bars represent three independent experiments and each experiment was triplicated. Means with the same letter are not significantly different by Duncan's multiple range test (P < 0.05). SJW: 70% ethanol extract of St. John's Wort

• 

  Fig. 3 Effect of St. John's Wort extract on the gene expression of alkaline phosphatase (A), osteocalcin (B) and osteoprotegerin (C) in MG 63 cells. Values are mean ± SD. Stand error bars represent three independent experiments and each experiment was triplicated. Means with the same letter are not significantly different by Duncan's multiple range test (P < 0.05). SJW: 70% ethanol extract of St. John's Wort

• 

  Fig. 4 Effect of St. John's Wort extract on ER α (A) and ER β (B) in MG 63 cells. Values are mean ± SD. Stand error bars represent three independent experiments and each experiment was triplicated. Means with the same letter are not significantly different by Duncan's multiple range test (P < 0.05). SJW: 70% ethanol extract of St. John's Wort

• 

  Fig. 5 Histopathology of the metaphyseal region of distal rat femur in each group (H&E). 100× magnification, scale bar = 200 µm. Sham-C: Sham operation + distilled water, OVX-C: Ovariectomy + distilled water, E: Ovariectomy + estradiol 50 µg/kg body weight, SJW100: Ovariectomy + St. John's Wort extract 100 mg/kg body weight, SJW200: Ovariectomy + St. John's Wort extract 200 mg/kg body weight

• 

  Fig. 6 Effect of St. John's Wort extract on Bone volume/tissue volume (A), Trabecular thickness (B), Trabecular number (C) and Trabecular separation (D) in ovariectomized rats. Values are mean ± SD. Means with the same letter are not significantly different by Duncan's multiple range test (P < 0.05). Sham-C: Sham operation + distilled water, OVX-C: Ovariectomy + distilled water, E: Ovariectomy + estradiol 50 µg/kg body weight, SJW100: Ovariectomy + St. John's Wort extract 100 mg/kg body weight, SJW200: Ovariectomy + St. John's Wort extract 200 mg/kg body weight

• 

  Fig. 7 Effect of St. John's Wort extract on the serum alkaline phosphatase (A) and C-telopeptide (B) Values are mean ± SD. Means with the same letter are not significantly different by Duncan's multiple range test (P < 0.05). Sham-C: Sham operation + distilled water, OVX-C: Ovariectomy + distilled water, E: Ovariectomy + estradiol 50 µg/kg body weight, SJW100: Ovariectomy + St. John's Wort extract 100 mg/kg body weight, SJW200: Ovariectomy + St. John's Wort extract 200 mg/kg body weight

Reference

1. Clowes JA, Riggs BL, Khosla S. The role of the immune system in the pathophysiology of osteoporosis. Immunol Rev. 2005; 208:207–227.

2. Marshall D, Johnell O, Wedel H. Meta-analysis of how well measures of bone mineral density predict occurrence of osteoporotic fractures. BMJ. 1996; 312:1254–1259.

3. Osteoporosis . review of the evidence for prevention, diagnosis and treatment and cost-effective analysis. Osteoporos Int. 1998; 8:Suppl 4. S1–88.

4. Royal College of Physicians of London. Osteoporosis: Clinical Guidelines for Prevention and Treatment. London: Royal College of Physicians of London;1999.

5. Lerner UH. Bone remodeling in post-menopausal osteoporosis. J Dent Res. 2006; 85:584–595.

6. Kousteni S, Han L, Chen JR, Almeida M, Plotkin LI, Bellido T, Manolagas SC. Kinase-mediated regulation of common transcription factors accounts for the bone-protective effects of sex steroids. J Clin Invest. 2003; 111:1651–1664.

7. Gallo D, Zannoni GF, Apollonio P, Martinelli E, Ferlini C, Passetti G, Riva A, Morazzoni P, Bombardelli E, Scambia G. Characterization of the pharmacologic profile of a standardized soy extract in the ovariectomized rat model of menopause: effects on bone, uterus, and lipid profile. Menopause. 2005; 12:589–600.

8. Mennini T, Gobbi M. The antidepressant mechanism of Hypericum perforatum. Life Sci. 2004; 75:1021–1027.

9. Brenner R, Azbel V, Madhusoodanan S, Pawlowska M. Comparison of an extract of hypericum (LI 160) and sertraline in the treatment of depression: a double-blind, randomized pilot study. Clin Ther. 2000; 22:411–419.

10. Schrader E. Equivalence of St John's wort extract (Ze 117) and fluoxetine: a randomized, controlled study in mild-moderate depression. Int Clin Psychopharmacol. 2000; 15:61–68.

11. You MK, Rhuy J, Jeong KS, Bang MA, Kim MS, Kim HA. Effect of St. John's Wort (Hypericum perforatum) on obesity, lipid metabolism and uterine epithelial proliferation in ovariectomized rats. Nutr Res Pract. 2014; 8:292–296.

12. Parfitt AM, Drezner MK, Glorieux FH, Kanis JA, Malluche H, Meunier PJ, Ott SM, Recker RR. Bone histomorphometry: standardization of nomenclature, symbols, and units. Report of the ASBMR Histomorphometry Nomenclature Committee. J Bone Miner Res. 1987; 2:595–610.

13. Dempster DW, Compston JE, Drezner MK, Glorieux FH, Kanis JA, Malluche H, Meunier PJ, Ott SM, Recker RR, Parfitt AM. Standardized nomenclature, symbols, and units for bone histomorphometry: a 2012 update of the report of the ASBMR Histomorphometry Nomenclature Committee. J Bone Miner Res. 2013; 28:2–17.

14. Cho YS, Jung WK, Kim JA, Choi IL, Kim SK. Beneficial effects of fucoidan on osteoblastic MG-63 cell differentiation. Food Chem. 2009; 116:990–994.

15. Reid P, Holen I. Pathophysiological roles of osteoprotegerin (OPG). Eur J Cell Biol. 2009; 88:1–17.

16. Udagawa N, Takahashi N, Yasuda H, Mizuno A, Itoh K, Ueno Y, Shinki T, Gillespie MT, Martin TJ, Higashio K, Suda T. Osteoprotegerin produced by osteoblasts is an important regulator in osteoclast development and function. Endocrinology. 2000; 141:3478–3484.

17. Dougall WC, Chaisson M. The RANK/RANKL/OPG triad in cancerinduced bone diseases. Cancer Metastasis Rev. 2006; 25:541–549.

18. Kim J, Um SJ, Woo J, Kim JY, Kim HA, Jang KH, Kang SA, Lim BO, Kang I, Choue RW, Cho Y. Comparative effect of seeds of Rhynchosia volubilis and soybean on MG-63 human osteoblastic cell proliferation and estrogenicity. Life Sci. 2005; 78:30–40.

19. Webb P, Nguyen P, Valentine C, Lopez GN, Kwok GR, McInerney E, Katzenellenbogen BS, Enmark E, Gustafsson JA, Nilsson S, Kushner PJ. The estrogen receptor enhances AP-1 activity by two distinct mechanisms with different requirements for receptor transactivation functions. Mol Endocrinol. 1999; 13:1672–1685.

20. Watkins MP, Norris JY, Grimston SK, Zhang X, Phipps RJ, Ebetino FH, Civitelli R. Bisphosphonates improve trabecular bone mass and normalize cortical thickness in ovariectomized, osteoblast connexin43 deficient mice. Bone. 2012; 51:787–794.

21. Hodgkinson A, Aaron JE, Horsman A, McLachlan MS, Nrodin BE. Effect of oophorectomy and calcium deprivation on bone mass in the rat. Clin Sci Mol Med. 1978; 54:439–446.

22. Wronski TJ, Lowry PL, Walsh CC, Ignaszewski LA. Skeletal alterations in ovariectomized rats. Calcif Tissue Int. 1985; 37:324–328.

23. Wronski TJ, Dann LM, Scott KS, Cintrón M. Long-term effects of ovariectomy and aging on the rat skeleton. Calcif Tissue Int. 1989; 45:360–366.

24. Faienza MF, Ventura A, Marzano F, Cavallo L. Postmenopausal osteoporosis: the role of immune system cells. Clin Dev Immunol. 2013; 2013:575936.

25. Srivastava AK, Bhattacharyya S, Castillo G, Miyakoshi N, Mohan S, Baylink DJ. Development and evaluation of C-telopeptide enzyme-linked immunoassay for measurement of bone resorption in mouse serum. Bone. 2000; 27:529–533.

26. Srivastava AK, Bhattacharyya S, Li X, Mohan S, Baylink DJ. Circadian and longitudinal variation of serum C-telopeptide, osteocalcin, and skeletal alkaline phosphatase in C3H/HeJ mice. Bone. 2001; 29:361–367.

27. Shan PF, Wu XP, Zhang H, Luo XH, Cao XZ, Xie H, Liu SP, Pi YZ, Fang TY, Liu H, Chen ZH, Zhong N, Liao EY. Age-related changes of serum bone alkaline phosphatase and cross-linked C-telopeptides of type I collagen and the relationship with bone mineral density in Chinese women. Clin Chim Acta. 2006; 366:233–238.