Tag Archives: animal

Ukrain No Benefit in Rats #2

Abstract

Intermittent three-month treatment with Ukrain in intact and ovariectomized rats. Part II: Effect on bone mineral density of the femur.

Ukrain, an acid alkaloid derivative of Chelidonium majus L., was administered intraperitoneally to ovariectomized and control sexually mature female rats at doses of 7, 14 and 28 mg/kg once daily for 10 days, followed by 10-day break. This procedure was repeated five times. At the end of the Ukrain treatment (24 h after the last dose of the drug) the right femora of the rats were harvested and the bone densitometric parameters of the whole bone and distal metaphysial and intertrochanteric subregions were assessed using the dual energy x-ray absorptiometry densitometric method. The results showed no apparent decrease in bone mineral density in groups of rats studied. A nearly significant (p = 0.08) decrease of bone mineral content was observed in ovariectomized rats treated with 14 mg/kg of Ukrain.

Jabłoński M, Gorzelak M, Patyra M, Jagiełło-Wójtowicz E
Drugs Exp Clin Res 2000
PMID: 11345047

Berberine Promotes Osteoblasts in Mouse Cells

Abstract

Berberine promotes osteoblast differentiation by Runx2 activation with p38 MAPK.

Berberine (BBR) has been implicated in bone biology. Although BBR reduces osteoporosis by enhancing BMD and inhibiting osteoclast activity, the effects of BBR on osteoblasts during the process of osteogenesis have not been thoroughly studied. In osteoblastic cells, BBR enhanced the expression of osteogenic marker genes including osteopontin and osteocalcin and promoted the transcriptional activity of the key osteogenic transcription factor Runx2. In osteoblasts, BBR increased the binding of Runx2 to the promoter region of osteopontin. The recruitment of co-factors such as p300 and HDAC1 to the promoter regions of osteopontin and osteocalcin was regulated by BBR, resulting in an enhancement in the expression of those genes. Furthermore, BBR activated p38 mitogen-activated protein kinase (MAPK) and increased cyclooxygenase 2 (COX2) expression, which are key factors in osteoblast differentiation. Consistently, a p38 MAPK-specific inhibitor attenuated the effect of BBR on osteogenesis, whereas p38 MAPK overexpression augmented BBR-induced osteogenic gene expression. Moreover, BBR stimulated bone area formation in calvarial organ culture. Taken together, these findings indicate that BBR promotes osteoblast differentiation through activation of Runx2 by p38 MAPK. Therefore, BBR may be a potential therapeutic agent to treat bone-related disorders including osteoporosis.

Lee HW, Suh JH, Kim HN, Kim AY…
J. Bone Miner. Res. Aug 2008
PMID: 18410224

Berberine Inhibits Osteoclasts in Mouse Cells

Abstract

Berberine inhibits RANKL-induced osteoclast formation and survival through suppressing the NF-kappaB and Akt pathways.

Berberine, an isoquinoline alkaloid isolated from several medicinal plants, has been reported to possess anti-bacterial, anti-inflammatory and antitumor properties. Although berberine also inhibits osteoclastogenesis and bone resorption, the molecular machinery for its inhibitory effects remains unknown. This study focused on the suppressive effects of berberine on receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL)-induced osteoclastogenesis and survival. Berberine inhibited RANKL-mediated osteoclast formation and survival while having no cytotoxic effects on bone marrow macrophages or osteoblastic cells. Berberine attenuated RANKL-induced activation of NF-kappaB through inhibiting phosphorylation at the activation loop of IkappaBalpha kinase beta, phosphorylation and degradation of IkappaBalpha, and NF-kappaB p65 nuclear translocation. RANKL-induced Akt phosphorylation was strongly inhibited by berberine; however, neither monocyte/macrophage-colony stimulating factor (M-CSF)-induced nor insulin-induced Akt activation was inhibited by the drug. Under M-CSF- and RANKL-deprived condition, berberine increased the active form of caspase-3 in osteoclasts. By contrast, berberine did not potentiate the activation of caspase-3 in M-CSF-deprived bone marrow macrophages. These findings indicate that berberine inhibits osteoclast formation and survival through suppression of NF-kappaB and Akt activation and that both pathways in the osteoclast lineage are highly sensitive to berberine treatment.

Hu JP, Nishishita K, Sakai E, Yoshida H…
Eur. J. Pharmacol. Feb 2008
PMID: 18083161

Berberine Increases Bone Density in Rats

Abstract

Effect of berberine on bone mineral density in SAMP6 as a senile osteoporosis model.

The effects of berberine in senescence accelerated mice P6 (SAMP6) were investigated to learn whether the alkaloid affects bone mineral density (BMD). Oral administration of berberine (10 mg/kg/d) to male and female mice for 22 weeks resulted in an increase in BMD in both sexes. A decreased concentration of deoxypyridinoline (Dpd) in urine was only observed in female mice. There was no effect on body or tibia weight or on the concentration of procollagen type I carboxyterminal extension peptide (PICP) in serum.

Li H, Miyahara T, Tezuka Y, Tran QL…
Biol. Pharm. Bull. Jan 2003
PMID: 12520186 | Free Full Text

Ukrain No Benefit in Rats

Abstract

Effect of intermittent three-month treatment with different doses of Ukrain on subregional femoral bone mineral density of sexually mature female rats.

Sexually mature but still growing female Wistar rats received i.p. injections of Ukrain (7, 14 or 28 mg/kg in a volume of 0.5 ml/100g) every other day for 10 days, followed by a 10-day break, and this procedure was performed five times. The control animals received the same volume of injected water. At the end of the experiment the rat right femora were harvested and the bone densitometric parameters of the entire bone, distal metaphyseal and basicervical subregions were assessed using the dual-energy X-ray absorptiometry (DXA) densitometric method. No significant changes were observed in the bone mineral density in experimental groups in comparison with control animals that received the vehicle. A slight decrease in the bone mineral content value was observed in the distal metaphyseal region in animals that were treated with the highest dose of Ukrain.

Gorzelak M, Jabłoński M, Patyra M, Jagiello-Wójtowicz E
Drugs Exp Clin Res 1998
PMID: 10190094

Ukrain Prevents Bone Loss in Rats

Abstract

Effect of six-month treatment with Ukrain on early osteoporosis induced by ovariectomy in rats. Part I: Preliminary studies of bone parameters.

Ukrain, thiophosphoric acid alkaloid derivatives from Chelidonium majus L. was administered intraperitoneally in a dose of 28 mg/kg (equivalent to 0.1 LD50) every other day for six months to female rats with ovariectomy-induced early osteoporosis. Administration of Ukrain was started on the second day after the surgical operation. At the end of the long-term treatment with Ukrain each rat was tested for the strength of both humeri and some parameters of rat femur were measured. The body weight of ovariectomized rats was also examined. The present results show that the decrease in the mechanical strength of the humeral bones and some changes in the femur caused by ovariectomy were prevented by the six-month treatment with Ukrain. However, in both ovariectomized groups and in ovariectomized rats pretreated with Ukrain an increase of body weight was observed.

Jagiełło-Wojtowicz E, Kleinrok Z, Nowicky JW, Jabłonski M…
Drugs Exp Clin Res 1996
PMID: 8899324

Vitamin C is a Skeletal Anabolic Agent in Mice

Abstract

Vitamin C prevents hypogonadal bone loss.

Epidemiologic studies correlate low vitamin C intake with bone loss. The genetic deletion of enzymes involved in de novo vitamin C synthesis in mice, likewise, causes severe osteoporosis. However, very few studies have evaluated a protective role of this dietary supplement on the skeleton. Here, we show that the ingestion of vitamin C prevents the low-turnover bone loss following ovariectomy in mice. We show that this prevention in areal bone mineral density and micro-CT parameters results from the stimulation of bone formation, demonstrable in vivo by histomorphometry, bone marker measurements, and quantitative PCR. Notably, the reductions in the bone formation rate, plasma osteocalcin levels, and ex vivo osteoblast gene expression 8 weeks post-ovariectomy are all returned to levels of sham-operated controls. The study establishes vitamin C as a skeletal anabolic agent.

Zhu LL, Cao J, Sun M, Yuen T…
PLoS ONE 2012
PMID: 23056580 | Free Full Text

Vitamin C Inhibits Osteoclasts in Rats Fed a High-Cholesterol Diet

Abstract

Vitamin C intake inhibits serum lipid peroxidation and osteoclast differentiation on alveolar bone in rats fed on a high-cholesterol diet.

A high-cholesterol diet stimulates osteoclast differentiation, which may be induced by increased serum lipid peroxidation. The inhibition of serum lipid peroxidation by vitamin C may offer beneficial effects on osteoclast differentiation including increased expression of receptor activator of nuclear factor (NF)-kappaB ligand (RANKL) and NF-kappaB. This study investigated the effects of vitamin C intake on RANKL and NF-kappaB expression in periodontal tissue of rats fed a high-cholesterol diet.
Twenty-four rats (8 weeks old) were divided into four groups: a control group (fed a regular diet) and three experimental groups (fed a high-cholesterol diet supplemented with 0, 1 and 2 g/l vitamin C/day) in this 12-week study. Vitamin C was provided by its addition to drinking water. As an index of serum lipid peroxidation, hexanoyl-lysine (HEL) level was determined by a competitive enzyme-linked immunosorbent assay method. Immunohistological analysis was performed to evaluate RANKL and NF-kappaB expression on the alveolar bone surface. The number of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts was also counted.
Feeding a high-cholesterol diet increased not only the serum HEL level but also the number of TRAP-positive osteoclasts on the alveolar bone surface, with an increase in RANKL and NF-kappaB expression on alveolar bone surface. Intake of vitamin C reduced the serum HEL level and osteoclast differentiation, with decreasing RANKL and NF-kappaB expression.
Vitamin C intake could suppress osteoclast differentiation, including RANKL and NF-kappaB expression on the alveolar bone surface, by decreasing serum lipid peroxidation in rats fed a high-cholesterol diet.

Sanbe T, Tomofuji T, Ekuni D, Azuma T…
Arch. Oral Biol. Mar 2009
PMID: 19110235

Vitamin C Reverses Bone Loss in Rats

Abstract

Evaluation of erythrocyte deformability in experimentally induced osteoporosis in female rats and the effects of vitamin C supplementation on erythrocyte deformability.

The aim of this study was to evaluate the possible variations in antioxidant enzymes, lipid peroxidation and erythrocyte deformability in experimentally induced osteoporosis in female rats and to assess the effects of vitamin C supplementation on those variations.
A total of 20 female Wistar Albino Rats were randomized into the three groups as controls (Group C, n = 6), ovariectomized rats (Group O, n = 7) and ovariectomized rats receiving vitamin C supplementation (Group OVC, n = 7). After the surgical procedure of ovariectomy, group OVC received 1 g ascorbic acid in 500 mL water daily. After 100 days following the ovariectomy, bone mineral density (BMD) was measured by using dual-energy X-ray absorptiometry.
BMD was significantly lower in the group O than in the group C (p = 0.015), whereas it was significantly higher in the group OVC than in the group O (p = 0.003). MDA activity was significantly higher in the group O than in the group C (p = 0.032), whereas it was significantly lower in the group OVC than in the group O (p = 0.025). SOD activity was significantly higher in the group O than in the group C (p = 0.032). Erythrocyte deformability was significantly higher in the group O than in the group C and OVC (p = 0.008, p = 0.021, respectively).
Erythrocyte deformability may show negative variations, suggesting a causative role in disruption of blood flow and tissue perfusion, which also negatively affect bone metabolism. Vitamin C supplementation seems to reverse those negative effects of variations in erythrocyte deformability. However, our preliminary results should be confirmed in more experimental studies and clinical trials (Tab. 3, Ref. 28).

Arslan A, Aydin G, Keles I, Fm C…
Bratisl Lek Listy 2011
PMID: 22180984

Lipoic Acid Helps Prevent Bone Destruction in Arthritic Mice

Abstract

Dietary alpha lipoic acid supplementation prevents synovial inflammation and bone destruction in collagen-induced arthritic mice.

Rheumatoid arthritis (RA) is a chronic inflammatory disorder characterized by chronic inflammation and joint destruction. In this study, we investigated whether dietary supplementation with alpha lipoic acid (ALA) suppresses collagen-induced arthritis (CIA) in mice. Mice were randomly divided into three groups: (1) a control CIA group was fed a normal diet, (2) a CIA group was fed a 0.1% ALA diet (average ALA intake of 160 mg/kg/day), and (3) a CIA group was fed a 0.5% ALA diet (average ALA intake of 800 mg/kg/day). The ALA-fed mice showed a decreased incidence and severity of arthritis compared to the normal diet group. Radiographic findings revealed a dramatic decrease in bone destruction, and histological findings showed extensively suppressed pathological changes in the ALA-fed mice. The ALA-fed mice exhibited inhibited generation of tartrate resistant acid phosphatase (TRAP)-positive osteoclasts in vivo. Additionally, ALA-fed mice reduced production of various proinflammatory cytokines and the soluble receptor activator of NF-κB ligand (sRANKL) in the joint tissues and the sera. In conclusion, dietary supplementation with ALA attenuated inflammatory responses and bone destruction in CIA mice.

Hah YS, Sung MJ, Lim HS, Jun JS…
Rheumatol. Int. Dec 2011
PMID: 20496068