Tag Archives: abstract

p-Hydroxycinnamic Acid Stimulates Mineralization in Mouse Osteoblastic Cells

Abstract

Phytocomponent p-hydroxycinnamic acid stimulates mineralization in osteoblastic MC3T3-E1 cells.

Phytocomponent p-hydroxycinnamic acid (HCA) has been shown to have stimulatory effects on bone calcification and inhibitory effects on bone resorption in rat femoral tissues in vitro. Whether HCA has a stimulatory effect on mineralization in osteoblastic cells is unknown. This study was undertaken to determine the effect of HCA on mineralization in osteoblastic MC3T3-E1 cells in vitro.Cells were cultured for 72 h in a minimum essential medium (alpha-MEM) containing 10% fetal bovine serum (FBS), and the cells with subconfluency were changed to a medium containing either vehicle or HCA (10(-7)-10(-5) M) without FBS. Culture with HCA (10(-7)-10(-5) M) did not have a significant effect on cell proliferation and cell death. Deoxyribonucleic acid (DNA) content in osteoblastic cells was significantly increased after culture with HCA (10(-6) or 10(-5) M) for 48 or 72 h. Alkaline phosphatase activity in osteoblastic cells was significantly increased after culture with HCA (10(-7)-10(-5) M) for 24, 48, or 72 h. The results with Alizarin red staining for calcium showed that mineralization was significantly stimulated after culture with HCA (10(-8)-10(-5) M) for 7, 14, or 21 days. This study demonstrates that HCA has stimulatory effects on mineralization in osteoblastic MC3T3-E1 cells.

Yamaguchi M, Lai YL, Uchiyama S, Nakagawa T
Int. J. Mol. Med. Sep 2008
PMID: 18698486

p-Hydroxycinnamic Acid Prevents Bone Loss in Diabetic Rats

Abstract

Oral administration of phytocomponent p-hydroxycinnamic acid has a preventive effect on bone loss in streptozotocin-induced diabetic rats.

The phytocomponent p-hydroxycinnamic acid (HCA) has been shown to have a stimulatory effect on bone formation and an inhibitory effect on bone resorption in rat femoral tissues in vitro. The preventive effect of HCA on bone loss induced in streptozotocin (STZ)-diabetic rats was investigated in vivo. Rats received a single subcutaneous administration of STZ (6.0 mg/100 g body weight), and then the animals were orally administered HCA (0.25, 0.5, or 1.0 mg/100 g body weight) once daily for 14 days. STZ administration caused a significant decrease in body weight and a significant increase in serum glucose, triglyceride, and calcium levels, indicating a diabetic state. These alterations were significantly prevented by administration of HCA (0.25, 0.5, or 1.0 mg/100 g). Calcium content in the femoral-diaphyseal and -metaphyseal tissues was significantly decreased in STZ-diabetic rats. This decrease was significantly prevented after administration of HCA (0.25, 0.5, or 1.0 mg/100 g). Alkaline phosphatase activity in the diaphyseal and metaphyseal tissues was significantly decreased in STZ-diabetic rats. The decrease in diaphyseal alkaline phosphatase activity in STZ-diabetic rats was significantly prevented after administration of HCA (0.5 and 1.0 mg/l00 g). The diaphyseal DNA content was also significantly decreased in STZ-diabetic rats. Administration of HCA (0.25, 0.5, or 1.0 mg/100 g) caused a significant increase in DNA content in the diaphyseal and metaphyseal tissues in STZ-diabetic rats. This study demonstrates that the intake of HCA has preventive effects on bone loss in STZ-diabetic rats, and that the intake has partially restorative effects on serum biochemical findings in the diabetic state.

Yamaguchi M, Uchiyama S, Lai YL
Int. J. Mol. Med. May 2007
PMID: 17390086

p-Hydroxycinnamic Acid Inhibits Osteoclast-Like Mouse Cells

Abstract

Phytocomponent p-hydroxycinnamic acid inhibits osteoclast-like cell formation in mouse bone marrow cultures.

The phytocomponent p-hydroxycinnamic acid (HCA) has been shown to have inhibitory effects on bone-resorbing factor-stimulated bone resorption in rat femoral tissues in vitro. The effects of HCA on osteoclast-like cell formation in mouse bone marrow cultures in vitro were investigated. The bone marrow cells were cultured for 7 days in alpha-minimal essential medium containing a bone-resorbing agent [parathyroid hormone (1-34)] (PTH), prostaglandin E2 (PGE2), or tumor necrosis factor-alpha (TNF-alpha) in effective concentrations. Osteoclast-like cell formation was estimated by staining for tartrate-resistant acid phosphatase, a marker enzyme of osteoclasts. The presence of PTH (10(-7) M), PGE2 (10(-5) M), or TNF-alpha (10 ng/ml) induced a remarkable increase in osteoclast-like multinucleated cells. These increases were significantly inhibited in the presence of HCA (10(-8)-10(-5) M). HCA (10(-6) or 10(-5) M) significantly inhibited osteoclast-like cell formation induced by dibutyryl cyclic adenosine monophosphate (10(-5) M) or phorbol 12-myristate 13-acetate (10(-6) M), an activator of protein kinase C. Also, HCA (10(-8)-10(-5) M) had a significant inhibitory effect on osteoclast-like cell formation induced by the receptor activator of NF-kappaB ligand (RANKL) (10 ng/ml) in the presence of macrophage colony-stimulating factor (M-CSF) (10 ng/ml). The inhibitory effect of HCA (10(-6) or 10(-5) M) on RANKL plus M-CSF-induced osteoclast-like cell formation was not observed in the presence of cycloheximide (10(-7) M), an inhibitor of protein synthesis in the transcriptional process, or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (10(-6) M), an inhibitor of transcription. This study demonstrates that HCA has a potent inhibitory effect on osteoclast-like cell formation in mouse bone marrow cultures. The inhibitory action of HCA may partly involve a newly synthesized protein component which is related to RANKL stimulation in osteoclastogenesis.

Lai YL, Yamaguchi M
Int. J. Mol. Med. Jan 2007
PMID: 17143556

Review: Phytochemicals for Bone Osteoporosis

Abstract

Regulatory mechanism of food factors in bone metabolism and prevention of osteoporosis.

Aging induces a decrease in bone mass, and osteoporosis with its accompanying decrease in bone mass is widely recognized as a major public health problem. Bone loss with increasing age may be due to decreased bone formation and increased bone resorption. Pharmacologic and nutritional factors may prevent bone loss with aging, although chemical compounds in food and plants which act on bone metabolism are poorly understood. We have found that isoflavones (including genistein and daidzein), which are contained in soybeans, have a stimulatory effect on osteoblastic bone formation and an inhibitory effect on osteoclastic bone resorption, thereby increasing bone mass. Menaquinone-7, an analogue of vitamin K(2) which is abundant in fermented soybeans, has been demonstrated to stimulate osteoblastic bone formation and to inhibit osteoclastic bone resorption. Of various carotenoids, beta-cryptoxanthin, which is abundant in Satsuma mandarin (Citrus unchiu MARC), has a stimulatory effect on osteoblastic bone formation and an inhibitory effect on osteoclastic bone resorption. The supplementation of these factors has a preventive effect on bone loss induced by ovariectomy in rats, which are an animal model of osteoporosis, and their intake has been shown to have a stimulatory effect on bone mass in humans. Factors with an anabolic effect on bone metabolism were found in extracts obtained from wasabi leafstalk (Wasabi japonica MATSUM), the marine alga Sargassum horneri, and bee pollen Cistus ladaniferus. Phytocomponent p-hydroxycinnamic acid was also found to have an anabolic effect on bone metabolism. Food chemical factors thus play a role in bone health and may be important in the prevention of bone loss with increasing age.

Yamaguchi M
Yakugaku Zasshi Nov 2006
PMID: 17077614 | Free Full Text

p-Hydroxycinnamic Acid Anabolic and Antiresorptive in Rats

Abstract

Phytocomponent p-hydroxycinnamic acid stimulates bone formation and inhibits bone resorption in rat femoral tissues in vitro.

The effect of cinnamic acid or its related compounds, which is present in many plants, on bone metabolism has not been clarified yet. The effect of cinnamic acid, p-hydroxycinnamic acid (HCA), ferulic acid, caffeic acid, or 3,4-dimethoxycinnamic acid (DCA) on bone calcium content in vitro was investigated. Rat femoral-diaphyseal (cortical bone) and -metaphyseal (trabecular bone) tissues were cultured for 48,h in Dulbecco’s modified Eagle’s medium (high glucose, 4.5%) supplemented with antibiotics and bovine serum albumin. The presence of HCA (10(-5) or 10(-4),M) caused a significant increase in calcium content in the diaphyseal or metaphyseal tissues. Such an effect was not observed in the presence of cinnamic acid or other compounds at the concentration of 10(-5) or 10(-4),M. Alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the diaphyseal or metaphyseal tissues was significantly increased in the presence of HCA (10(-5) or 10(-4),M). The effect of HCA (10(-4),M) in increasing calcium content, alkaline phosphatase activity, and DNA content in the diaphyseal or metaphyseal tissues was completely prevented in the presence of cycloheximide (10(-6),M), an inhibitor of protein synthesis. Thus HCA had anabolic effects on bone components. The presence of parathyroid hormone (PTH; 10(-7),M), a bone-resorbing factor, caused a significant decrease in calcium content and a corresponding elevation in medium glucose consumption, lactic acid production or tartrate-resistant acid phosphatase (TRACP) activity in the diaphyseal or metaphyseal tissues. These alterations were completely prevented in the presence of HCA (10(-5) or 10(-4),M). This study demonstrates that p-hydroxycinnamic acid (HCA) has stimulatory effects on bone formation and inhibitory effects on bone resorption in tissue culture in vitro.

Lai YL, Yamaguchi M
Mol. Cell. Biochem. Nov 2006
PMID: 17036165

p-Hydroxycinnamic Promotes Osteoblasts by Blocking NF-κB in Mouse Cells

Abstract

The bone anabolic carotenoid p-hydroxycinnamic acid promotes osteoblast mineralization and suppresses osteoclast differentiation by antagonizing NF-κB activation.

Numerous plant derived nutritional factors including p-hydroxycinnamic acid (HCA), a member of the carotenoid family, have long been held to possess bone protective properties. Studies in animals have provided a mechanistic basis for these observations by demonstrating the capacity of HCA to promote bone formation and suppress bone resorption in vivo. However, the molecular mechanism by which HCA achieves these effects remains unclear. We have demonstrated that a centralized mechanism by which several other nutritional factors achieve similar effects is through modulation of the nuclear factor-κB (NF-κB) signal transduction pathway. NF-κB activation is essential for osteoclast formation and resorption but potently antagonizes osteoblast differentiation and mineralization. In this study we demonstrate that HCA does indeed antagonize the activation of NF-κB by the key osteoclastogenic cytokine receptor activator of NF-κB (RANKL) in RAW264.7 osteoclast precursors, suppressing their differentiation into osteoclasts. Furthermore, HCA augmented the in vitro differentiation of MC3T3 preosteoblastic cells into mineralizing osteoblasts and relieved the inhibitory action of tumor necrosis factor-α (TNF-α)-induced NF-κB signaling on transforming growth factor-β (TGF-β)- or bone morphogenetic protein-2 (BMP-2)-induced Smad activation, an important pathway in osteoblast commitment and differentiation. Our data provide a mechanism to explain the dual pro-anabolic and anti-catabolic activities of HCA.

Yamaguchi M, Weitzmann MN
Int. J. Mol. Med. Sep 2012
PMID: 22751682

p-Hydroxycinnamic Acid Prevents Bone Loss in Ovariectomized Rats

Abstract

Oral administration of phytocomponent p-hydroxycinnamic acid prevents bone loss in ovariectomized rats.

The preventive effect of phytocomponent p-hydroxycinnamic acid (HCA) on ovariectomy (OVX)-induced bone loss was investigated. HCA (250 or 500 microg/100 g body weight) was orally administered once daily for 30 days to OVX rats. The analysis using a peripheral quantitative computed tomography (pQCT) showed that OVX caused bone loss in the femoral-metaphyseal tissues. This change was significantly restored after the administration of HCA (250 or 500 microg/100 g body weight) to OVX rats. Mineral content, mineral density, and polar strength strain index in the femoral-metaphyseal tissues were significantly decreased in OVX rats. These decreases were significantly restored after the administration of HCA (500 microg/100 g) to OVX rats. Moreover, OVX caused a significant decrease in calcium content or alkaline phosphatase activity in the femoral-diaphyseal and -metaphyseal tissues. These decreases were significantly restored after the administration of HCA (250 or 500 microg/100 g) to OVX rats. Deoxyribonucleic acid (DNA) content in the diaphyseal or metaphyseal tissues was significantly increased in OVX rats. These increases were significantly restored after oral administration of HCA (500 microg/100 g). This study demonstrates that HCA has preventive effects on OVX-induced bone loss of rats in vivo.

Yamaguchi M, Lai YL, Uchiyama S, Nakagawa T
Mol. Cell. Biochem. Apr 2008
PMID: 18165927

Mountain Tea Protects Bone in Ovariectomized Rats

Abstract

Protective effect of Sideritis euboea extract on bone mineral density and strength of ovariectomized rats.

The aim of this study was to investigate the potential protective effect of Sideritis euboea extract (SID), commonly consumed as “mountain tea,” on bone mineral density (BMD) and the strength of the ovariectomized (OVX) rat model of osteoporosis.
Thirty-two 10-month-old Wistar rats were separated into controls (sham operated), OVX, and OVX plus SID in their drinking water (dose, 330 mg/kg body weight per day), starting immediately after OVX for 6 months. Tibial BMD at baseline and at 3 and 6 months post-OVX, three-point-bending of the femur, and body and uterine weight at the study end were examined.
BMD percentage change from baseline of the whole tibia was similar in control and OVX + SID rats at 3 months (-3.02% vs -4.67%, P = not significant), revealing a strong osteoprotective effect. At 6 months, the corresponding changes were -6.02% versus -14.37%, P < 0.05, indicating a greater bone loss in treated rats, albeit significantly less than the OVX change (-20.46%; OVX vs OVX + SID, P < 0.05). The proximal (metaphyseal) tibial BMD percentage change from baseline to 3 and 6 months between the OVX and OVX + SID groups (-26.47% vs -15.57% and -31.22% vs -16.57%, respectively) was statistically significant, demonstrating that SID preserved the proximal tibial BMD of the OVX + SID group significantly. Three-point-bending showed a significant increase in the treated compared with the OVX groups. Body and uterine weights were similar in the OVX and treated groups.
SID significantly protected tibial bone loss and improved femoral biomechanical strength in OVX + SID rats compared with OVX rats.

Dontas IA, Lelovas PP, Kourkoulis SK, Aligiannis N…
Menopause Aug 2011
PMID: 21505372

Minocycline Increases Bone Formation and Decreases Resorption in Ovariectomized Rats

Abstract

Minocycline prevents the decrease in bone mineral density and trabecular bone in ovariectomized aged rats.

In the current study, we examined the effects of minocycline, on the osteopenia of ovariectomized aged rats. Old female rats were randomly divided into five groups: sham, ovariectomized control and ovariectomized treated with minocycline, 17beta-estradiol, or both agents. Bone samples were collected 8 wk after the treatment. Ovariectomy reduced bone mineral density of the whole femur and at the condylar, distal metaphyseal and head-neck-trochanter regions 10%-19% and the loss of bone density was prevented by treatment with minocycline or 17beta-estradiol. Histomorphometric analysis of distal femur showed ovariectomy reduced the trabecular bone area, the trabecular bone number, trabecular bone thickness and increased the trabecular bone separation. The microanatomic structure of trabecular bone also showed that the number of nodes, node to node, cortical to node, node to free end was reduced by ovariectomy. Treatment with minocycline attenuated the effect of ovariectomy on trabecular bone in aged animals. In contrast, cortical bone was not affected by ovariectomy or minocycline treatment. The effect of minocycline on bone turnover was also examined. Minocycline increased osteoid surface, mineralizing surface, mineral apposition rate, bone formation rate and reduced eroded surface. We have therefore concluded that the modest increase in bone mineral density and the improvement in the trabecular bone status noted in minocycline treated ovariectomized aged rats is likely due to an increase in bone formation coupled with a decrease in bone resorption.

Williams S, Wakisaka A, Zeng QQ, Barnes J…
Bone Dec 1996
PMID: 8968031

Tetracyclines Prevent Bone Loss Induced by Inflammation

Abstract

Tetracyclines convert the osteoclastic-differentiation pathway of progenitor cells to produce dendritic cell-like cells.

Tetracyclines, such as doxycycline and minocycline, are used to suppress the growth of bacteria in patients with inflammatory diseases. Tetracyclines have been shown to prevent bone loss, but the mechanism involved is unknown. Osteoclasts and dendritic cells (DCs) are derived from common progenitors, such as bone marrow-derived macrophages (BMMs). In this article, we show that tetracyclines convert the differentiation pathway, resulting in DC-like cells not osteoclasts. Doxycycline and minocycline inhibited the receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis of BMMs, but they had no effects on cell growth and phagocytic activity. They influenced neither the proliferation nor the differentiation of bone-forming osteoblasts. Surprisingly, doxycycline and minocycline induced the expression of DC markers, CD11c and CD86, in BMMs in the presence of RANKL. STAT5 is involved in DC differentiation induced by GM-CSF. Midostaurin, a STAT5-signaling inhibitor, and an anti-GM-CSF-neutralizing Ab suppressed the differentiation induced by GM-CSF but not by tetracyclines. In vivo, the injection of tetracyclines into RANKL-injected mice and RANKL-transgenic mice suppressed RANKL-induced osteoclastogenesis and promoted the concomitant appearance of CD11c(+) cells. These results suggested that tetracyclines prevent bone loss induced by local inflammation, including rheumatoid arthritis and periodontitis, through osteoclast-DC-like cell conversion.

Kinugawa S, Koide M, Kobayashi Y, Mizoguchi T…
J. Immunol. Feb 2012
PMID: 22250082 | Free Full Text