Prevention of Osteoporosis in the Ovariectomized Rat by Oral Administration of a Nutraceutical Combination That Stimulates Nitric Oxide Production

Osteoporosis represents an imbalance between bone formation and bone resorption. As a result of low estrogen levels, it is markedly prevalent during menopause, thus making such patients susceptible to fractures. Both bone formation and resorption are modulated by nitric oxide (NO). Currently, there...

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Main Authors: Rebecca A. Rajfer, Manuel Flores, Andrea Abraham, Eduardo Garcia, Natalhy Hinojosa, Mina Desai, Jorge N. Artaza, Monica G. Ferrini
Format: Article
Language:English
Published: Wiley 2019-01-01
Series:Journal of Osteoporosis
Online Access:http://dx.doi.org/10.1155/2019/1592328
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author Rebecca A. Rajfer
Manuel Flores
Andrea Abraham
Eduardo Garcia
Natalhy Hinojosa
Mina Desai
Jorge N. Artaza
Monica G. Ferrini
author_facet Rebecca A. Rajfer
Manuel Flores
Andrea Abraham
Eduardo Garcia
Natalhy Hinojosa
Mina Desai
Jorge N. Artaza
Monica G. Ferrini
author_sort Rebecca A. Rajfer
collection DOAJ
description Osteoporosis represents an imbalance between bone formation and bone resorption. As a result of low estrogen levels, it is markedly prevalent during menopause, thus making such patients susceptible to fractures. Both bone formation and resorption are modulated by nitric oxide (NO). Currently, there are no risk-free pharmaceutical prevention therapies for osteoporosis. COMB-4, a nutraceutical combination of Paullinia cupana, Muira puama, ginger, and L-citrulline, known to activate the NO-cGMP pathway, was reported to accelerate fracture healing in the rat. To determine whether COMB-4 could be effective in preventing menopausal osteoporosis, it was compared to estradiol (E2) in an ovariectomized (OVX) rat osteoporosis model. Nine-month-old female Sprague Dawley rats were divided into SHAM, OVX, OVX+E2, and OVX+COMB-4. After 100 days of treatment, bone mineral density (BMD) and bone mineral content (BMC) were measured by DXA scan. TRAP staining was performed in the femur and lumbar vertebrae. TRACP 5b and osteocalcin levels were assayed in the serum. MC3T3-E1 cells were differentiated into osteoblasts and treated with COMB-4 for one week in order to evaluate calcium deposition by Alizarin staining, cGMP production by ELISA, and upregulation of the nitric oxide synthase (NOS) enzymes by RT-PCR. OVX resulted in a decrease in BMD, BMC, and serum osteocalcin and an increase in serum TRACP 5b. Except for an increase in BMC with COMB-4, both E2 and COMB-4 reverted all bone and serum markers, as well as the number of osteoclasts in the vertebrae, to SHAM levels. Incubation of MC3T3-E1 cells with COMB-4 demonstrated an increase in the three NOS isoforms, cGMP, and calcium deposition. COMB-4 increased BMD in OVX rats by inhibiting bone resorption and increasing calcium deposition presumably via activation of the NO-cGMP pathway. It remains to be determined whether COMB-4 could be a potential nutraceutical therapy for the prevention of premenopausal bone loss.
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spelling doaj-art-aede6b35909a4498bf78f93b96efc93c2025-08-20T03:39:00ZengWileyJournal of Osteoporosis2090-80592042-00642019-01-01201910.1155/2019/15923281592328Prevention of Osteoporosis in the Ovariectomized Rat by Oral Administration of a Nutraceutical Combination That Stimulates Nitric Oxide ProductionRebecca A. Rajfer0Manuel Flores1Andrea Abraham2Eduardo Garcia3Natalhy Hinojosa4Mina Desai5Jorge N. Artaza6Monica G. Ferrini7Department of Orthopedic Surgery, Loma Linda University, Loma Linda, CA, USADepartment of Health and Life Sciences, Charles R. Drew University of Medicine and Science, Los Angeles, CA, USADepartment of Health and Life Sciences, Charles R. Drew University of Medicine and Science, Los Angeles, CA, USADepartment of Health and Life Sciences, Charles R. Drew University of Medicine and Science, Los Angeles, CA, USADepartment of Health and Life Sciences, Charles R. Drew University of Medicine and Science, Los Angeles, CA, USAPerinatal Research Laboratory, Department of Obstetrics and Gynecology, Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, Torrance, CA, USADepartment of Health and Life Sciences, Charles R. Drew University of Medicine and Science, Los Angeles, CA, USADepartment of Health and Life Sciences, Charles R. Drew University of Medicine and Science, Los Angeles, CA, USAOsteoporosis represents an imbalance between bone formation and bone resorption. As a result of low estrogen levels, it is markedly prevalent during menopause, thus making such patients susceptible to fractures. Both bone formation and resorption are modulated by nitric oxide (NO). Currently, there are no risk-free pharmaceutical prevention therapies for osteoporosis. COMB-4, a nutraceutical combination of Paullinia cupana, Muira puama, ginger, and L-citrulline, known to activate the NO-cGMP pathway, was reported to accelerate fracture healing in the rat. To determine whether COMB-4 could be effective in preventing menopausal osteoporosis, it was compared to estradiol (E2) in an ovariectomized (OVX) rat osteoporosis model. Nine-month-old female Sprague Dawley rats were divided into SHAM, OVX, OVX+E2, and OVX+COMB-4. After 100 days of treatment, bone mineral density (BMD) and bone mineral content (BMC) were measured by DXA scan. TRAP staining was performed in the femur and lumbar vertebrae. TRACP 5b and osteocalcin levels were assayed in the serum. MC3T3-E1 cells were differentiated into osteoblasts and treated with COMB-4 for one week in order to evaluate calcium deposition by Alizarin staining, cGMP production by ELISA, and upregulation of the nitric oxide synthase (NOS) enzymes by RT-PCR. OVX resulted in a decrease in BMD, BMC, and serum osteocalcin and an increase in serum TRACP 5b. Except for an increase in BMC with COMB-4, both E2 and COMB-4 reverted all bone and serum markers, as well as the number of osteoclasts in the vertebrae, to SHAM levels. Incubation of MC3T3-E1 cells with COMB-4 demonstrated an increase in the three NOS isoforms, cGMP, and calcium deposition. COMB-4 increased BMD in OVX rats by inhibiting bone resorption and increasing calcium deposition presumably via activation of the NO-cGMP pathway. It remains to be determined whether COMB-4 could be a potential nutraceutical therapy for the prevention of premenopausal bone loss.http://dx.doi.org/10.1155/2019/1592328
spellingShingle Rebecca A. Rajfer
Manuel Flores
Andrea Abraham
Eduardo Garcia
Natalhy Hinojosa
Mina Desai
Jorge N. Artaza
Monica G. Ferrini
Prevention of Osteoporosis in the Ovariectomized Rat by Oral Administration of a Nutraceutical Combination That Stimulates Nitric Oxide Production
Journal of Osteoporosis
title Prevention of Osteoporosis in the Ovariectomized Rat by Oral Administration of a Nutraceutical Combination That Stimulates Nitric Oxide Production
title_full Prevention of Osteoporosis in the Ovariectomized Rat by Oral Administration of a Nutraceutical Combination That Stimulates Nitric Oxide Production
title_fullStr Prevention of Osteoporosis in the Ovariectomized Rat by Oral Administration of a Nutraceutical Combination That Stimulates Nitric Oxide Production
title_full_unstemmed Prevention of Osteoporosis in the Ovariectomized Rat by Oral Administration of a Nutraceutical Combination That Stimulates Nitric Oxide Production
title_short Prevention of Osteoporosis in the Ovariectomized Rat by Oral Administration of a Nutraceutical Combination That Stimulates Nitric Oxide Production
title_sort prevention of osteoporosis in the ovariectomized rat by oral administration of a nutraceutical combination that stimulates nitric oxide production
url http://dx.doi.org/10.1155/2019/1592328
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