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Pomegranate Peel Extract Prevents Bone Loss in a Preclinical Model of Osteoporosis and Stimulates Osteoblastic Differentiation in Vitro.

Spilmont M, Léotoing L, Davicco MJ, Lebecque P, Miot-Noirault E, Pilet P, Rios L, Wittrant Y, Coxam V - Nutrients (2015)

Bottom Line: The nutritional benefits of pomegranate have attracted great scientific interest.Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia.In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP) activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers.

View Article: PubMed Central - PubMed

Affiliation: Unité de Nutrition Humaine, CRNH Auvergne, UMR 1019, INRA, F-63000 Clermont-Ferrand, France. mel.spilmont@gmail.com.

ABSTRACT
The nutritional benefits of pomegranate have attracted great scientific interest. The pomegranate, including the pomegranate peel, has been used worldwide for many years as a fruit with medicinal activity, mostly antioxidant properties. Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia. In this study, with in vivo and ex vivo experiments, we investigated whether the consumption of pomegranate peel extract (PGPE) could limit the process of osteopenia. We demonstrated that in ovariectomized (OVX) C57BL/6J mice, PGPE consumption was able to significantly prevent the decrease in bone mineral density (-31.9%; p < 0.001 vs. OVX mice) and bone microarchitecture impairment. Moreover, the exposure of RAW264.7 cells to serum harvested from mice that had been given a PGPE-enriched diet elicited reduced osteoclast differentiation and bone resorption, as shown by the inhibition of the major osteoclast markers. In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP) activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers. PGPE may be effective in preventing the bone loss associated with ovariectomy in mice, and offers a promising alternative for the nutritional management of this disease.

No MeSH data available.


Related in: MedlinePlus

Expression profile analysis of bone (A), oxidative stress and inflammation (B) markers in femoral bone from mice fed the control diet (Sham-operated, SH Control. Ovariectomized, OVX Control) or exposed to pomegranate peel extract (OVX PGPE, diet containing 2 g/kg of PGPE), for 30 days. Transcriptomic analysis of bone tissue mRNA levels determined by Taqman Low density Arrays are presented as fold change compared to OVX Control group (fold change = 1). Results are expressed as mean ± standard deviation, SD (n = 8). * p < 0.05. (A) Osteoclast genes: CTR: calcitonin receptor; Fos; ITG b3: integrin β3; MMP2: metalloproteinase 2. Osteoblast genes: Lrp5; OCN: osteocalcin; OPN: osteopontin; RunX2. (B) Oxidative stress genes: GPx: glutathione peroxidase; GSR: glutathione reductase; NOS2: nitric oxide synthase 2; NOX4: NADPH oxidase 4. Inflammation genes: CCL2: Chemokine (C-C motif) ligand 2; INFγR1: interferon gamma receptor 1; IL1-R1: interleukin 1 receptor 1; IL1-Rn: interleukin 1 receptor antagonist.
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nutrients-07-05465-f005: Expression profile analysis of bone (A), oxidative stress and inflammation (B) markers in femoral bone from mice fed the control diet (Sham-operated, SH Control. Ovariectomized, OVX Control) or exposed to pomegranate peel extract (OVX PGPE, diet containing 2 g/kg of PGPE), for 30 days. Transcriptomic analysis of bone tissue mRNA levels determined by Taqman Low density Arrays are presented as fold change compared to OVX Control group (fold change = 1). Results are expressed as mean ± standard deviation, SD (n = 8). * p < 0.05. (A) Osteoclast genes: CTR: calcitonin receptor; Fos; ITG b3: integrin β3; MMP2: metalloproteinase 2. Osteoblast genes: Lrp5; OCN: osteocalcin; OPN: osteopontin; RunX2. (B) Oxidative stress genes: GPx: glutathione peroxidase; GSR: glutathione reductase; NOS2: nitric oxide synthase 2; NOX4: NADPH oxidase 4. Inflammation genes: CCL2: Chemokine (C-C motif) ligand 2; INFγR1: interferon gamma receptor 1; IL1-R1: interleukin 1 receptor 1; IL1-Rn: interleukin 1 receptor antagonist.

Mentions: To elucidate the possible molecular mechanisms involved in this effect on bone morphology and to confirm the in vitro findings, we performed a transcriptomic analysis on bone tissue samples using Taqman Low Density Arrays (TLDA, Applied Biosystems, Life Technologies, Saint Aubin, France). Because of the large number of genes modulated, only the most important bone markers are presented. Data on the OVX PGPE group were compared to the data measured for the OVX Control group. The TLDA data on osteoclasts are shown in Figure 5A. Accordingly, PGPE consumption significantly (p < 0.05) down-regulated the mRNA levels of some of the major osteoclast-specific markers: define (0.39 fold) and integrin β3 (ITG b3: 0.56 fold). Fos, a crucial transcription factor for osteoclast differentiation, was also inhibited in the PGPE group (0.22 fold; p < 0.05). Finally, we observed a trend toward down-regulated expression in the OVX PGPE group of a key enzyme for osteoclast activity involved in collagen degradation: the metallo-proteinase 2 (MMP2) (PGt: 0.55; not significant). In addition, regarding the expression of osteoblast-specific markers, the TLDA data presented in Figure 5A show that PGPE consumption by ovariectomized mice was able to significantly improve the expression of a co-receptor implied in Wnt/β-catenin signalling in the osteoblast, LRP5 (1.62 fold; p < 0.05), in comparison with the OVX Control group. However, the other osteoblast markers investigated were not significantly modulated by dietary PGPE.


Pomegranate Peel Extract Prevents Bone Loss in a Preclinical Model of Osteoporosis and Stimulates Osteoblastic Differentiation in Vitro.

Spilmont M, Léotoing L, Davicco MJ, Lebecque P, Miot-Noirault E, Pilet P, Rios L, Wittrant Y, Coxam V - Nutrients (2015)

Expression profile analysis of bone (A), oxidative stress and inflammation (B) markers in femoral bone from mice fed the control diet (Sham-operated, SH Control. Ovariectomized, OVX Control) or exposed to pomegranate peel extract (OVX PGPE, diet containing 2 g/kg of PGPE), for 30 days. Transcriptomic analysis of bone tissue mRNA levels determined by Taqman Low density Arrays are presented as fold change compared to OVX Control group (fold change = 1). Results are expressed as mean ± standard deviation, SD (n = 8). * p < 0.05. (A) Osteoclast genes: CTR: calcitonin receptor; Fos; ITG b3: integrin β3; MMP2: metalloproteinase 2. Osteoblast genes: Lrp5; OCN: osteocalcin; OPN: osteopontin; RunX2. (B) Oxidative stress genes: GPx: glutathione peroxidase; GSR: glutathione reductase; NOS2: nitric oxide synthase 2; NOX4: NADPH oxidase 4. Inflammation genes: CCL2: Chemokine (C-C motif) ligand 2; INFγR1: interferon gamma receptor 1; IL1-R1: interleukin 1 receptor 1; IL1-Rn: interleukin 1 receptor antagonist.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4663593&req=5

nutrients-07-05465-f005: Expression profile analysis of bone (A), oxidative stress and inflammation (B) markers in femoral bone from mice fed the control diet (Sham-operated, SH Control. Ovariectomized, OVX Control) or exposed to pomegranate peel extract (OVX PGPE, diet containing 2 g/kg of PGPE), for 30 days. Transcriptomic analysis of bone tissue mRNA levels determined by Taqman Low density Arrays are presented as fold change compared to OVX Control group (fold change = 1). Results are expressed as mean ± standard deviation, SD (n = 8). * p < 0.05. (A) Osteoclast genes: CTR: calcitonin receptor; Fos; ITG b3: integrin β3; MMP2: metalloproteinase 2. Osteoblast genes: Lrp5; OCN: osteocalcin; OPN: osteopontin; RunX2. (B) Oxidative stress genes: GPx: glutathione peroxidase; GSR: glutathione reductase; NOS2: nitric oxide synthase 2; NOX4: NADPH oxidase 4. Inflammation genes: CCL2: Chemokine (C-C motif) ligand 2; INFγR1: interferon gamma receptor 1; IL1-R1: interleukin 1 receptor 1; IL1-Rn: interleukin 1 receptor antagonist.
Mentions: To elucidate the possible molecular mechanisms involved in this effect on bone morphology and to confirm the in vitro findings, we performed a transcriptomic analysis on bone tissue samples using Taqman Low Density Arrays (TLDA, Applied Biosystems, Life Technologies, Saint Aubin, France). Because of the large number of genes modulated, only the most important bone markers are presented. Data on the OVX PGPE group were compared to the data measured for the OVX Control group. The TLDA data on osteoclasts are shown in Figure 5A. Accordingly, PGPE consumption significantly (p < 0.05) down-regulated the mRNA levels of some of the major osteoclast-specific markers: define (0.39 fold) and integrin β3 (ITG b3: 0.56 fold). Fos, a crucial transcription factor for osteoclast differentiation, was also inhibited in the PGPE group (0.22 fold; p < 0.05). Finally, we observed a trend toward down-regulated expression in the OVX PGPE group of a key enzyme for osteoclast activity involved in collagen degradation: the metallo-proteinase 2 (MMP2) (PGt: 0.55; not significant). In addition, regarding the expression of osteoblast-specific markers, the TLDA data presented in Figure 5A show that PGPE consumption by ovariectomized mice was able to significantly improve the expression of a co-receptor implied in Wnt/β-catenin signalling in the osteoblast, LRP5 (1.62 fold; p < 0.05), in comparison with the OVX Control group. However, the other osteoblast markers investigated were not significantly modulated by dietary PGPE.

Bottom Line: The nutritional benefits of pomegranate have attracted great scientific interest.Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia.In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP) activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers.

View Article: PubMed Central - PubMed

Affiliation: Unité de Nutrition Humaine, CRNH Auvergne, UMR 1019, INRA, F-63000 Clermont-Ferrand, France. mel.spilmont@gmail.com.

ABSTRACT
The nutritional benefits of pomegranate have attracted great scientific interest. The pomegranate, including the pomegranate peel, has been used worldwide for many years as a fruit with medicinal activity, mostly antioxidant properties. Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia. In this study, with in vivo and ex vivo experiments, we investigated whether the consumption of pomegranate peel extract (PGPE) could limit the process of osteopenia. We demonstrated that in ovariectomized (OVX) C57BL/6J mice, PGPE consumption was able to significantly prevent the decrease in bone mineral density (-31.9%; p < 0.001 vs. OVX mice) and bone microarchitecture impairment. Moreover, the exposure of RAW264.7 cells to serum harvested from mice that had been given a PGPE-enriched diet elicited reduced osteoclast differentiation and bone resorption, as shown by the inhibition of the major osteoclast markers. In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP) activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers. PGPE may be effective in preventing the bone loss associated with ovariectomy in mice, and offers a promising alternative for the nutritional management of this disease.

No MeSH data available.


Related in: MedlinePlus