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Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (10): 1125-1132.doi: 10.3736/jcim20111014

• Original Experimental Research • Previous Articles     Next Articles

Anti-osteoporotic activity of methanolic extract of an Indian herbal formula NR/CAL/06 in ovariectomized rats

Praveen Srikanta1, Shivaprasad H. Nagarajappa2, Gollapalle L. Viswanatha1(), Mukund Handral1, Rajesh Subbanna1, Rangappa Srinath1, Ganesh Hiremath1#br#   

  1. 1. Department of Pharmacology, People’s Education Society (PES) College of Pharmacy, Bangalore 560050, India
    2. Natural Remedies Pvt. Ltd, Veerasandra Industrial Area, Bangalore 560100, India
  • Received:2011-06-24 Accepted:2011-08-05 Online:2011-10-20 Published:2011-10-15
  • Contact: L. Viswanatha Gollapalle E-mail:glv_000@yahoo.com

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Objective: The present study was aimed to evaluate the anti-osteoporotic activity of methanolic extract of NR/CAL/06 in ovariectomized (OVX) rats.

Methods: Bilateral ovariectomy was performed in female Sprague-Drawley rats under aseptic conditions and the rats were divided into five groups (n=10). Two different doses of methanolic extract of NR/CAL/06 (200 and 400 mg/kg, per oral (p.o.)) were evaluated for anti-osteoporotic activity and raloxifene (5.4 mg/kg, p.o.) was used as a reference standard. Treatment was given for 90 d. Anti-osteoporotic potential of NR/CAL/06 was evaluated based on various parameters, namely, body weight, organ weight, bone weight, bone mineral content, bone strength, calcium and phosphorus in serum and urine, and serum alkaline phosphatase (ALP). External diameter, length and thickness of the femur bone were measured by using scanning electron microscopy (SEM).

Results: The bilateral ovariectomy in rats resulted in decreased bone strength, bone mineral content and bone weight. The SEM images showed porous, perforated and disintegrated femur bone architecture and decreases in bone length and thickness in OVX rats. These changes were associated with elevated serum levels of calcium, phosphorus and ALP. Increases in body weight and adipose weight and a decrease in uterine weight were also observed and the changes were highly significant when compared with the sham-control group. Treatment with methanolic extract of NR/CAL/06 (200 and 400 mg/kg, p.o.) for 90 d dose-dependently restored the ovariectomy-induced alterations in bone weight, bone mineral content, bone strength, serum calcium, phosphorus and ALP, body weight and adipose weight nearly to normal levels. Furthermore, the SEM images of the femur bones of NR/CAL/06 (200 and 400 mg/kg, p.o.)-treated rats showed reduced pore formation and improved bone compactness compared with the OVX-control group.

Conclusion: These findings suggest that the methanolic extract of NR/CAL/06 possesses significant anti-osteoporotic activity and may be useful in the treatment of osteoporosis.

Key words: osteoporosis, ovariectomy, plant extracts, medicine, Ayurvedic, microscopy, electron, scanning, rats

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Group n Body weight (g) Increase in body
weight (%)
Day 0 Day 45 Day 90
Sham-control 10 198.20±3.43 209.10±6.87 232.60±6.71 17.47
OVX-control 10 213.10±9.26 233.70±8.18 272.00±9.77** 28.12**
Raloxifene (5.4 mg/kg) 10 209.60±13.19 222.29±14.42 244.15±14.45△△ 16.82△△
NR/CAL/06 (200 mg/kg) 10 216.87±38.63 229.90±41.90 258.24±44.67 19.56△△
NR/CAL/06 (400 mg/kg) 10 208.80±5.38 226.45±4.77 242.33±5.39 16.50△△

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Group n Adipose mass
(g)		 Uterine weight
(g)		 Femur weight
(g)		 Femur length
(mm)		 Femur thickness
(mm)
Sham-control 10 2.642±0.775 0.383±0.014 0.713±0.021 35.028±0.760 3.907±0.120
OVX-control 10 3.857±0.747** 0.173±0.043** 0.690±0.178* 35.597±9.007 3.792±0.949*
Raloxifene (5.4 mg/kg) 10 1.907±0.394△△ 0.223±0.035△△ 0.808±0.166△△ 35.853±7.310 3.905±0.797△
NR/CAL/06 (200 mg/kg) 10 3.107±0.608△ 0.220±0.051△△ 0.810±0.168△△ 35.797±6.926 3.862±0.752△
NR/CAL/06 (400 mg/kg) 10 2.703±0.394△△ 0.297±0.095△△ 0.818±0.170△△ 36.210±7.562 3.933±0.823△

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Group n Calcium (mg/dL) Phosphorus (mg/dL)
Day 0 Day 45 Day 90 Day 0 Day 45 Day 90
Sham-control 10 10.08±0.19 10.13±0.13 10.09±0.09 3.04±0.34 2.70±0.17 3.05±0.18
OVX-control 10 10.09±0.17 11.08±0.05** 11.21±0.07** 3.95±0.37 4.08±0.16 5.97±0.10**
Raloxifene (5.4 mg/kg) 10 9.98±0.11 10.25±0.05△△ 10.23±0.07△△ 3.02±0.08 2.47±0.14 2.62±0.08△△
NR/CAL/06 (200 mg/kg) 10 10.16±0.14 10.25±0.09△ 10.15±0.07△△ 4.18± 0.51 2.87±0.39 2.52±0.43△△
NR/CAL/06 (400 mg/kg) 10 10.29±0.13 10.18±0.07△ 10.21±0.08△△ 3.02±0.27 2.94±0.07 3.19±0.12△△
Group n ALP (Iu/L)
Day 0 Day 45 Day 90
Sham-control 10 110.9±14.0 119.2±14.6 128.4±14.0
OVX-control 10 111.9±17.2 262.8±23.7** 318.9±39.6**
Raloxifene (5.4 mg/kg) 10 119.7±11.3 165.3±25.3 155.6±27.2△△
NR/CAL/06 (200 mg/kg) 10 121.4±18.0 158.0±25.0 149.9±24.4△△
NR/CAL/06 (400 mg/kg) 10 113.5±07.9 148.2±24.3 147.9±29.8△△

"

Group n Urine calcium
(mg/dL)		 Urine phosphorus
(mg/dL)		 Bone mineral content
Ash weight (g) Ash calcium
(mg/g Ash)		 Ash phosphorus
(mg/g Ash)
Sham-control 10 4.860±0.534 9.860±0.634 0.650±0.034 11.6±0.12 6.50±0.54
OVX-control 10 8.190±0.374** 11.200±0.774* 0.580±0.054** 10.0±0.14** 6.56±0.56
Raloxifene (5.4 mg/kg) 10 4.920±0.088△△ 7.840±0.688△ 0.670±0.059△△ 10.8±0.13△△ 6.56±0.56
NR/CAL/06 (200 mg/kg) 10 6.350±0.513△△ 8.750±0.813 0.600±0.055 11.3±0.11△△ 6.47±0.60
NR/CAL/06 (400 mg/kg) 10 6.160±0.274△ 8.610±0.674 0.620±0.053△ 11.9±0.10△△ 6.53±0.53

"

Group n Force (N)
Sham-control 10 112.800±4.885
OVX-control 10 78.540±2.974**
Raloxifene (5.4 mg/kg) 10 168.000±5.948△△
NR/CAL/06 (200 mg/kg) 10 74.270±2.182
NR/CAL/06 (400 mg/kg) 10 99.170±2.148△△

Figure 1

Bone architecture of rats in different groups observed under a scanning electron microscopeMagnifications are 1 000×. Arrow indicates the pore formation and reduced compactness. The OVX-control group showed porous perforated and disintegrated bone architecture compared with the sham-control group. NR/CAL/06 (200 mg/kg) treatment showed partial restoration; NR/CAL/06 (400 mg/kg)- and raloxifene (5.4 mg/kg)-treated groups showed complete restoration of osteoporosis bone changes to a compact firm.A: Sham-control; B: OVX-control; C: NR/CAL/06 (200 mg/kg); D NR/CAL/06 (400 mg/kg); E: Raloxifene (5.4 mg/kg). OVX: ovariectomized."

[1] Canalis E, Giustina A, Bilezikian JP . Mechanisms of anabolic therapies for osteoporosis[J]. N Engl J Med, 2007,357(9):905-916
doi: 10.1056/NEJMra067395 pmid: 17761594
[2] Heaney RP, Recker RR, Saville PD . Menopausal changes in bone remodeling[J]. J Lab Clin Med, 1978,92(6):964-970
pmid: 739174
[3] McNamara LM, Ederveen AG, Lyons CG, Price C, Schaffler MB, Weinans H, Prendergast PJ . Strength of cancellous bone trabecular tissue from normal, ovariectomized and drug-treated rats over the course of ageing[J]. Bone, 2006,39(2):392-400
doi: 10.1016/j.bone.2006.02.070
[4] Basly JP, Lavier MC . Dietary phytoestrogens: potential selective estrogen enzyme modulators?[J]. Planta Med, 2005,71(4):287-294
doi: 10.1055/s-2005-864092 pmid: 15856402
[5] Mundy GR . Osteoporosis into the year 2010[J]. Br J Obstet Gynaecol, 1996,103(Suppl 13):32-37
pmid: 8624340
[6] Somekawa Y, Wakabayashi A . Relationship between apolipoprotein E polymorphism, menopausal symptoms, and serum lipids during hormone replacement therapy[J]. Eur J Obstet Gynecol Reprod Biol, 1998,79(2):185-191
doi: 10.1016/S0301-2115(98)00008-6
[7] Gambacciani M, Ciaponi M . Postmenopausal osteoporosis management[J]. Curr Opin Obstet Gynecol, 2000,12(3):189-197
doi: 10.1097/00001703-200006000-00005
[8] Stēpán JJ, Musilová J, Pacovský V . Bone demineralization, biochemical indices of bone remodeling, and estrogen replacement therapy in adults with Turner’s syndrome[J]. J Bone Miner Res, 1989,4(2):193-198
[9] Goldstein SR, Siddhanti S, Ciaccia AV, Plouffe L Jr . A pharmacological review of selective oestrogen receptor modulators[J]. Hum Reprod Update, 2000,6(3):212-224
doi: 10.1093/humupd/6.3.212 pmid: 10874566
[10] Gorman C, Park A . The truth about hormones[J]. Time, 2002,160(4):32-39
[11] Vasudeva N, Sharma SK . Post-coital antifertility activity of Hibiscus rosasinensis Linn. roots[J]. Evid Based Complement Alternat Med, 2008,5(1):91-94
doi: 10.1093/ecam/nem003 pmid: 18317554
[12] Ghosh A, Chowdhury N, Chandra G . Laboratory evaluation of a phytosteroid compound of mature leaves of Day Jasmine(Solanaceae: Solanales) against larvae of Culex quinquefasciatus(Diptera: Culicidae) and nontarget organisms[J]. Parasitol Res, 2008,103(2):271-277
doi: 10.1007/s00436-008-0963-y
[13] Obolentseva GV, Litvinenko VI, Ammosov AS, Popova TP, Sampiev AM . Pharmacological and therapeutic properties of licorice preparations(A review)[J]. Pharm Chem J, 1999,33(8):24-31
doi: 10.1007/BF02508412
[14] Vaya J, Belinky PA, Aviram M . Antioxidant constituents from licorice roots: isolation, structure elucidation and antioxidative capacity toward LDL oxidation[J]. Free Radic Biol Med, 1997,23(2):302-313
doi: 10.1016/S0891-5849(97)00089-0
[15] Tamir S, Eizenberg M, Somjen D, Izrael S, Vaya J . Estrogen-like activity of glabrene and other constituents isolated from licorice root[J]. J Steroid Biochem Mol Biol, 2001,78(3):291-298
doi: 10.1016/S0960-0760(01)00093-0 pmid: 11595510
[16] Coxam V . New advances in osteoporosis nutritional prevention[J]. Med Sci (Paris), 2005,21(3):297-301
doi: 10.1051/medsci/2005213297
[17] Fujioka M, Uehara M, Wu J, Adlercreutz H, Suzuki K, Kanazawa K, Takeda K, Yamada K, Ishimi Y . Equol, a metabolite of daidzein, inhibits bone loss in ovariectomized mice[J]. J Nutr, 2004,134(10):2623-2627
doi: 10.1111/j.1365-277X.2004.00554.x pmid: 15465757
[18] Morin S . Isoflavones and bone health[J]. Menopause, 2004,11(3):239-241
doi: 10.1097/01.GME.0000125650.82154.B5
[19] Ke HZ, Chen HK, Simmons HA, Qi H, Crawford DT, Pirie CM, Chidsey-Frink KL, Ma YF, Jee WS, Thompson DD . Comparative effects of droloxifene, tamoxifen, and estrogen on bone, serum cholesterol, and uterine histology in the ovariectomized rat model[J]. Bone, 1997,20(1):31-39
doi: 10.1016/S8756-3282(96)00313-4
[20] Evans DB, Bunning RA, Russell RG . The effects of recombinant human interleukin-1β on cellular proliferation and the production of prostaglandin E2, plasminogen activator, osteocalcin and alkaline phosphatase by osteoblast-like cells derived from human bone[J]. Biochem Biophys Res Commun, 1990,166(1):208-216
doi: 10.1016/0006-291X(90)91932-I
[21] Einhorn TA . Bone strength: the bottom line[J]. Calcif Tissue Int, 1992,51(5):333-339
doi: 10.1007/BF00316875
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