Abstract
As the average age of society increases, identifying and preventing osteoporosis becomes
more important. According to the results of the Women’s Health Initiative study, substitution
of estradiol is not recommended in hormone replacement therapy (HRT), although phytoestrogens
might be a safe alternative. In this study, the osteoprotective effects of genistein
(Gen), resveratrol (Res) and 8-prenylnaringenin (8PN) were evaluated by analysing
bone biomechanical strength and bone mineral density. After ovariectomy, 88 female
rats received soy-free food (C), and according to their grouping, were fed estradiol
(E), GEN, RES or 8PN for 12 weeks. The phytohormones were given in two dosages. To
analyse the osteoprotective effects of the tested substances, bone biomechanical properties
and bone mineral density (BMD) were evaluated on the upper tibial metaphysis. Bone
biomechanical properties were significantly improved after treatment with E (Fmax : 90.6 N) and 8PN (85.0 N) compared to GEN (76.0 N), RES (72.6 N) and C (76.6 N).
Bone biomechanical properties with 8PN (yL: 55.7 N) supplementation reached a level
similar to that seen after E (49.3 N) supplementation. Treatment with GEN (38.5 N)
was not as effective as E and 8PN, but demonstrated improved biomechanical properties
compared to C (40.1 N) and RES (36.3 N). E (Cn.Dn. 217 mg/cm3 ) and 8PN (165 mg/cm3) showed superior results in the analysis of bone mineral density
compared to C (112 mg/cm3 ). GEN (164 mg/cm3 ) also demonstrated superior results, though not as good as E and 8PN. RES (124 mg/cm3 ) revealed no effect on bone density. Treatment with 8PN resulted in very good biomechanical
properties and showed an increased BMD. GEN had a smaller effect on bone biomechanical
strength, while RES did not have an effect on bone biomechanical strength or BMD.
Therefore, 8PN might be a safe alternative for HRT, but further studies are needed.
Abbreviations
BMD:total bone mineral density
C:control
Cn.Dn.:cancellous bone density
Ct.Dn.:cortical bone density
E:estradiol
fL:failure load
Fmax :maximum load
GEN:genistein
HRT:hormone replacement therapy
8PN:8-prenylnaringenin
RES:resveratrol
S:stiffness
yL:yield load
Key words
osteoporosis - biomechanical testing - 8-prenylnaringenin - genistein - resveratrol
References
1
Pacifici R.
Cytokines, estrogen, and postmenopausal osteoporosis – the second decade.
Endocrinology.
1998;
139
2659-61
2
Wronski T J, Yen C F.
The ovariectomized rat as an animal model for postmenopausal bone loss.
Cells Mater Supp.
1991;
11
69-74
3
Thompson D D, Simmons H A, Pirie C M, Ke H Z.
FDA Guidelines and animal models for osteoporosis.
Bone.
1995;
17
125-33
4
Fuse H, Fukumoto S, Sone H, Miyata Y, Saito T, Nakayama K. et al .
A new synthetic steroid, osaterone acetate (TZP-4238), increases cortical bone mass
and strength by enhancing bone formation in ovariectomized rats.
J Bone Miner Res.
2003;
12
590-7
5
Sturmer E K, Seidlova-Wuttke D, Sehmisch S, Rack T, Wille J, Frosch K H. et al .
Standardized bending and breaking test for the normal and osteoporotic metaphyseal
tibias of the rat: effect of estradiol, testosterone, and raloxifene.
J Bone Miner Res.
2006;
21
89-96
6
Rossouw J E, Anderson G L, Prentice R L, LaCroix A Z, Kooperberg C, Stefanick M L.
et al .
Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal
results From the Women′s Health Initiative randomized controlled trial.
JAMA 2002;.
17;
288
321-33
7
Bennets H W, Underwood E J, Shier F L.
A specific breeding problem of sheep on subterranean clover pastures in western Australia.
Aust Vet J.
1946;
22
2-12.
8
Holmes P, Rumsby P, Harrison P T.
Endocrine disrupters and menopausal health.
J Br Menopause Soc.
2004;
10
54-9
9
Li W, Seifert M, Xu Y, Hock B.
Comparative study of estrogenic potencies of estradiol, tamoxifen, bisphenol-A and
resveratrol with two in vitro bioassays.
Environ Int.
2004;
30
329-35
10
Sirtori C R.
Risks and benefits of soy phytoestrogens in cardiovascular diseases, cancer, climacteric
symptoms and osteoporosis.
Drug Saf.
2001;
24
665-82
11
Humpel M, Isaksson P, Schaefer O, Kaufmann U, Ciana P, Maggi A. et al .
Tissue specificity of 8-prenylnaringenin: protection from ovariectomy induced bone
loss with minimal trophic effects on the uterus.
J Steroid Biochem Mol Biol.
2005;
97
299-305
12
Hertrampf T, Degen G H, Kaid A A, Laudenbach-Leschowsky U, Seibel J, Di Virgilio A L.
et al .
Combined effects of physical activity, dietary isoflavones and 17β-estradiol on movement
drive, body weight and bone mineral density in ovariectomized female rats.
Planta Med.
2006;
72
84-7
13
Yamagishi T, Otsuka E, Hagiwara H.
Reciprocal control of expression of mRNAs for osteoclast differentiation factor and
OPG in osteogenic stromal cells by genistein: evidence for the involvement of topoisomerase
II in osteoclastogenesis.
Endocrinology.
2001;
142
3632-7
14
Rickard D J, Monroe D G, Ruesink T J, Khosla S, Riggs B L, Spelsberg T C.
Phytoestrogen genistein acts as an estrogen agonist on human osteoblastic cells through
estrogen receptors alpha and beta.
J Cell Biochem.
2003;
89
633-46
15
Christoffel J, Rimoldi G, Wuttke W.
Effects of 8-prenylnaringenin on the hypothalamo-pituitary-uterine axis in rats after
3-month treatment.
J Endocrinol.
2006;
188
397-405
16
Mesiano S, Katz S L, Lee J Y, Jaffe R B.
Phytoestrogens alter adrenocortical function: genistein and daidzein suppress glucocorticoid
and stimulate androgen production by cultured adrenal cortical cells.
J Clin Endocrinol Metab.
1999;
84
2443-8
17
Alarcon C, Villegas I.
Resveratrol as an anti-inflammatory and anti-aging agent: Mechanisms and clinical
implications.
Mol Nutr Food Res.
2005;
49
405-30
18
Bottner M, Christoffel J, Jarry H, Wuttke W.
Effects of long-term treatment with resveratrol and subcutaneous and oral estradiol
administration on pituitary function in rats.
J Endocrinol.
2006;
189
77-88
19
Barkhem T, Carlsson B, Nilsson Y, Enmark E, Gustafsson J, Nilsson S.
Differential response of estrogen receptor alpha and estrogen receptor beta to partial
estrogen agonists/antagonists.
Mol Pharmacol.
1998;
54
105-12
20
Zierau O, Gester S, Schwab P, Metz P, Kolba S, Wulf M. et al .
Estrogenic activity of the phytoestrogens naringenin, 6-(1,1-dimethylallyl)naringenin
an 8-prenylnaringenin.
Planta Med.
2002;
68
49-51
21
Goldstein S A.
The mechanical properties of trabecular bone: Dependence on anatomic location and
function.
J Biomech.
1994;
20
1055-61
22
Verhas M, Schoutens A, L′Hermite-Baleriaux M, Dourov N, Verschaeren A, Mone M. et
al .
The effect of orchidectomy on bone metabolism in aging rats.
Calcif Tissue Int.
1986;
39
74-7
23
Wronski T J, Schenck P A, Cintron M, Walsh C C.
Effect of body weight on osteopenia in ovariectomized rats.
Calcif Tissue Int.
1987;
40
155-9
24
Takeda S, Elefteriou F, Levasseur R, Liu X, Zhao L, Parker K L. et al .
Leptin regulates bone formation via the sympathetic nervous system.
Cell.
2002;
111
305-17
25
Roudebush R E, Magee D E, Benslay D N, Bendele A M, Bryant H U.
Effect of weight manipulation on bone loss due to ovariectomy and the protective effects
of estrogen in the rat.
Calcif Tissue Int.
1993;
53
61-4
26
Rad M, Humpel M, Schaefer O, Schoemaker R C, Schleuning W D, Cohen A F. et al .
Pharmacokinetics and systemic endocrine effects of the phyto-oestrogen 8-prenylnaringenin
after single oral doses to postmenopausal women.
Br J Clin Pharmacol.
2006;
62
288-96
27
Anderson J J, Ambrose W W, Garner S C.
Biphasic effects of genistein on bone tissue in the ovariectomized, lactating rat
model.
Proc Soc Exp Biol Med.
1998;
217
345-50
28
Alekel D L, Germain A S, Peterson C T, Hanson K B, Stewart J W, Toda T.
Isoflavone-rich soy protein isolate attenuates bone loss in the lumbar spine of perimenopausal
women.
Am J Clin Nutr.
2000;
72
844-52
29
Basly J P, Canivenc Lavier M C.
Dietary phytoestrogens: potential selective estrogen enzyme modulators?.
Planta Med.
2005;
71
87-94
30
Russell R M, Rasmussen H, Lichtenstein A H.
Modified food guide pyramid for people over seventy years of age.
J Nutr.
1999;
129
751-3
31
Mizutani K, Ikeda K, Kawai Y, Yamori Y.
Resveratrol stimulates the proliferation and differentiation of osteoblastic MC3T3-E1
cells.
Biochem Biophys Res Commun.
1998;
30;253
859-63
32
Supornsilchai V, Svechnikov K, Seidlova-Wuttke D, Wuttke W, Soder O.
Phytoestrogen resveratrol suppresses steroidogenesis by rat adrenocortical cells by
inhibiting cytochrome P450 c21-hydroxylase.
Horm Res.
2005;
64
280-6
Dr. Stephan Sehmisch
Department of Trauma and Reconstructive Surgery
University of Goettingen
Robert Koch-Str. 40
37075 Goettingen
Germany
Telefon: +49-551-39-2462
Fax: +49-551-39-8981
eMail: stephan.sehmisch@med.uni-goettingen.de