The Effects of Pueraria lobata on Osteoclast Differentiation and Bone Resorption / 체질인류학회지

Previous researches have proved that Pueraria lobata up-regulates bone mineral contents and bone mineral density in bone-loss model, ovariectomized mice and orchidectomized rats. However, the precise effects and mechanisms of Pueraria lobata on osteoclast differentiation and bone resorbing activity of mature osteoclasts still remains unknown. Therefore, we investigated the effect and mechanism of Pueraria lobata on receptor activator of nuclear factor-kappaB ligand (RANKL) and macrophage colony stimulation factor (M-CSF)-induced osteoclast differentiation in bone marrow macro-phages (BMMs). First of all, we treated BMMs derived from mice with various concentrations of Pueraria lobata in order to perform screening by tartrate-resistant acid phosphatase (TRAP) staining. Also, we conducted western blotting and RT-PCR for the purpose of verifying the treatment mechanism of Pueraria lobata and lastly, we used hydroxyapatite-coated plate to evaluate the effects of Pueraria lobata on bone resorbing activity of mature osteoclasts. As a result, Pueraria lobata has inhibitory effect on phosphorylation of p38, Akt, c-Jun N-terminal kinase (JNK), and IkappaB which are essential early signaling pathway of osteoclastogenesis. Also, the inactivation of nuclear factor of activated T cells (NFAT)c1, and c-Fos which is caused by Pueraria lobata is followed by the suppression effects of Pueraria lobata on osteoclast-related various genes, osteoclast-associated receptor (OSCAR), TRAP, Integrin beta3, osteoclast stimulatory transmembrane protein (OC-STAMP), and dendritic cell-specific transmembrane protein (DC-STAMP). Particularly, Pueraria lobata blocks the formation of pit area on hydroxyapatite-coated plate in a dose-dependent manner as well as the mRNA expression of Cathepsin K, which is associated with bone resorbing activity. These results demonstrate the molecular mechanism relating to anti-osteoclastogenesis effect of Pueraria lobata as well as the inhibitory effect of Pueraria lobata on mature osteoclast formation and bone resorbing activity.

The Effects of Baicalein on Osteoclast Differentiation from Bone Marrow Derived Macrophage / 체질인류학회지

As prediction of rapidly aging society, bone health is considered increasingly important and received more attention than ever. Bone health is regulated by balancing between bone resorptive osteoclasts and bone formative osteoblasts. Disruption of balance between bone-resorbing osteoclasts and bone-forming osteoblasts results in bone disease. Natural products have recently received much attention as an alternative tool for the development of novel therapeutic strategy. Baicalein is reported it has anti-cancer, anti-inflammatory and antioxidant effects. Baicalein also has been known that it has both promotive effect on MC3T3-E1 cell line and inhibitory effect on RAW 264.7 cell line. However, the inhibitory mechanism of baicalein using bone marrow derived macrophages (BMMs) on osteoclast differentiation remains not clear. In this study, the suppressive mechanism by baicalein on osteoblast differentiation was evaluated. Bicalein inhibited receptor activator of nuclear factor-kappaB ligand (RANKL)-induced osteoclast differentiation in BMMs in a dose dependent manner without any toxicity. Baicalein suppressed phosphorylation of protein kinaseB (Akt), c-Jun N-terminal kinases (JNK) and phosphoinositide-specific phospholipaseCgamma2 (PLCgamma2). Furthermore, Baicalein suppressed the induction of RANKL-induced c-Fos and Nuclear factor of activated T cell c1 (NFATc1), essential genes on osteoclastogenesis. In BMMs, Bicalein inhibited the mRNA expression of tartrate-resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), cathepsinK, dendritic cell-specific transmembrane protein (DC-STAMP). Moreover, baicalein promoted differentiation of osteoblast on bone marrow stromal cells (BMSCs). Taken together, these results suggest that baicalein has a potential for treating bone lytic diseases, such as osteoporosis, periodontitis, and rheumatoid arthritis.

The Effects of Phlomis umbrosa Turcz on Osteoclast Differentiation / 체질인류학회지

Many bone-related diseases such as osteoporosis, rheumatoid arthritis are occurred by excessive bone resorbing activity of osteoclasts. Recently, many studies have been proceeded to find out the new therapeutic materials from natural products of plants. Phlomis umbrosa Turcz, one of the natural products of plants has been known to improve bone health. However, the precise effects and treatment mechanisms of Phlomis umbrosa Turcz about bone diseases has been unknown. So, we examined the effects of Phlomis umbrosa Turcz on expression of receptor activator of nuclear factor-kappaB ligand (RANKL)-induced osteoclast differentiation in bone marrow-derived macrophages (BMMs) and bone resorption. Also, we investigated the treatment mechanisms of Phlomis umbrosa Turcz relating to osteoclast differentiation. Here, we showed that Phlomis umbrosa Turcz significantly suppressed RANKL-induced osteoclast differentiation and bone resportion. Furthermore, Phlomis umbrosa Turcz suppressed the activation of NF-kappaB in bone marrow macrophage treated RANKL and M-CSF. The mRNA expression of c-Fos, nuclear factor of activated T-cells (NFAT)c1, osteoclast-associated receptor (OSCAR), tartrate-resistant acid phosphatase (TRAP) in BMMs was inhibited by Phlomis umbrosa Turcz. Integrin alphanu, beta3 relating to cell adhesion and dendritic cell-specific transmembrane protein (DC-STAMP) relating to the structure of filamentous actin (F-actin) ring and cathepsin K relating to bone resorbing activity are disrupted too. These results suggest that Phlomis umbrosa Turcz will be a good materials to treat bone diseases like osteoporosis.

Effect of Cornus Officinalis on Receptor Activator of Nuclear Factor-kappaB Ligand (RANKL)-induced Osteoclast Differentiation

OBJECTIVES: Osteoporosis is a disease of bones that is thought to result from an imbalance between bone resorption and bone formation. Although osteoporosis itself has no symptoms, osteoporosis caused by osteoclasts leads to an increased risk of fracture. Here we examined the effects of cornus officinalis on receptor activator of nuclear factor-kappaB ligand (RANKL)-mediated osteoclast differentiation. METHODS: We evaluated the effects of cornus officinalis on RANKL-induced osteoclast differentiation from bone marrow-derived macrophages (BMMs) and performed a cytotoxicity assay, reverse transcriptase-polymerase chain reaction (RT-PCR), and Western blot analysis. RESULTS: Cornus officinalis significantly inhibits RANKL-mediated osteoclast differentiation in a dose-dependent manner, but without cytotoxicity against BMMs. The mRNA expression of tartrate-resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), c-Fos, and nuclear factor of activated T cells cytoplasmic 1 (NFATc1) in BMMs treated with RANKL was considerably inhibited by cornus officinalis treatment. Also, cornus officinalis inhibits the protein expression of c-Fos and NFATc1. Cornus officinalis greatly inhibits RANKL-induced phosphorylation of p38 and c-JUN N-terminal kinase (JNK). Also, cornus officinalis significantly suppresses RANKL-induced degradation of I-kappaB. CONCLUSIONS: Taken together, our results suggest that cornus officinalis may be a useful the treatment of osteoporosis.

The Respiratory Morbidities in Late-preterm Infants Compared with the Early-preterm and Term Infants throughout the First Year of Life

PURPOSE: We examined the respiratory morbidities in late-preterm infants compared to those of the early-preterm infants and term infants throughout the first year of life. METHODS: Data were retrospectively collected for 87 late-preterm, 72 early-preterm, and 608 term infants who were admitted to NICU and the nursery of Busan St. Mary's Medical Center from Jan 2007 to Oct 2009. RESULTS: There were significant differences in the proportions of the out-born infants, twin pregnancy, small for gestational age, and Caesarean section in the three groups (P<0.05). Late-preterm and early-preterm infants had longer duration of hospitalization, larger proportions of respiratory distress syndrome, mechanical ventilation at birth, oxygen therapy after 48 hours of birth, oxygen dependency at 28 days, and continuous positive airway pressure support at 28 days compared to term infants during the neonatal period (P=0.000). Late-preterm infants and early-preterm infants were re-admitted more often than term infants during the first year of life (P=0.000). Also Late-preterm and early-preterm infants had increased chance of respiratory tract illness than term infants (P=0.001). CONCLUSION: In this study, we demonstrated that there are higher chances of respiratory morbidities in the late-preterm infants than the term infants either during the neonatal period or throughout the first year of life, although early-preterm infants showed greatest respiratory morbidities.

Effects of Liriopis Tuber Water Extract on RANKL-induced Osteoclast Differentiation / 체질인류학회지

Increased formation and activation of osteoclast lead to unwanted bone resorption. Several natural products which have inhibitory effects on osteoclast differentiation and function are under investigation to prevent and treat the osteoporotic bone disease. Liriopis tuber has been used in Oriental medicine for the suppression of cough, expectoration, thirst, and has been used for sthenia, diuresis, blood glucose regulation, treatment of xerostomia, and constipation. Also, recently it has been reported that Liriopsis tuber has anti-inflammatory, anti-thrombic, anti-adhesive activities. The purpose of this study was to evaluate the effects of Liriopis tuber on osteoclast differentiation and was to evaluate of its mechanism. Water extract of Liriopis tuber significantly inhibited receptor activator of nuclear factor-kappaB ligand (RANKL)-induced osteoclast differentiation in bone marrow macrophages (BMMs) in a dose dependent manner. However, water extract of Liriopis tuber did not affect cytotoxicity when compared with control. The mRNA expression of c-Fos, NFATc1, tartrate resistant-acid phosphatase (TRAP), and cathepsin K induced by RANKL was inhibited by water extract of Liriopis tuber treatment. Also, water extract of Liriopis tuber inhibited the protein expression of c-Fos and NFATc1 expression in BMMs treated with RANKL. Among the signal pathways, water extract of Liriopis tuber suppressed the phosphorylation of p38 induced by RANKL. In summary, Liriopis tuber exerted inhibitory effects on osteoclast diffentiation via suppression of c-Fos and NFATc1 which are essential gene to osteoclastogenesis. Taken together, these results suggest that Liriopis tuber may be a useful candidate in the treatment of osteoporosis without special toxicity.

Inhibitory Effects of 1',2'-Dihydrorotenone on Osteoclast Differentiation and Bone Resorption In Vitro and In Vivo / 체질인류학회지

It is important to identify therapeutic compounds with no adverse effects for use in the chemotherapy of patients with bone-related diseases. The aim of this study was to identify a new compound that inhibits osteoclast differentiation and bone resorption. Herein, we examined the effects of 1',2'-dihydrorotenone on osteoclast differentiation and bone resorption in vitro and in vivo. 1',2'-dihydrorotenone inhibited receptor activator of NF-kappaB ligand (RANKL)-induced osteoclast differentiation of cultured bone marrow macrophages (BMMs) in a dose-dependent manner. However, 1',2'-dihydrorotenone did not exert cytotoxic effect on BMMs. 1',2'-dihydrorotenone suppressed the expression of c-fos and NFATc1 as well as osteoclast-specific genes in BMMs treated with RANKL. Treatment with RANKL inhibited the expression of inhibitors of differentiation/DNA binding (Id)1, 2, and 3; however, in the presence of 1',2'-dihydrorotenone, RANKL did not suppress the expression of Id1, 2, and 3. Furthermore, 1',2'-dihydrorotenone inhibited bone resorption and considerably attenuated the erosion of trabecular bone induced by lipopolysaccharide treatment. Taken together, these results suggest that 1',2'-dihydrorotenone has the potential to be applied in therapies for bone-related diseases.

Amorphigenin inhibits Osteoclast differentiation by suppressing c-Fos and nuclear factor of activated T cells / 대한해부학회지

Among the several rotenoids, amorphigenin is isolated from the leaves of Amopha Fruticosa and it is known that has anti-proliferative effects and anti-cnacer effects in many cell types. The main aim of this study was to investigate the effects of amorphigenin on osteoclast differentiation in vitro and on LPS treated inflammatory bone loss model in vivo. We show here that amorphigenin inhibited RANKL-induced osteoclast differentiation from bone marrow macrophages in a dose dependent manner without cellular toxicity. Anti-osteoclastogenic properties of amorphigenin were based on a down-regulation of c-fos and NFATc1. Amorphigenin markedly inhibited RANKL-induced p38 and NF-kappaB pathways, but other pathways were not affected. Micro-CT analysis of the femurs showed that amorphigenin protected the LPS-induced bone loss. We concluded that amorphigenin can prevent inflammation-induced bone loss. Thus we expect that amorphigenin could be a treatment option for bone erosion caused by inflammation.

Effect of Water Extract of Papaya on RANKL-induced Osteoclast Differentiation / 대한해부학회지

Balance between bone-resorbing osteoclats and bone-forming osteoblasts is important in bone homeostasis. In particular, increased osteoclast formation and activity are responsible for bone diseases such as osteoporosis, rheumatoid arthritis, periodontal disease. Natural metabolites of plants have recently received much attention as lead compounds for the development of novel therapeutic strategy. The purpose of this study was to search the natural products to inhibition osteoclast differentiation. Water extract of papaya significantly inhibited receptor activator of nuclear factor-kappaB ligand (RANKL)-induced osteoclast differentiation in bone marrow macrophages (BMMs) in a dose dependent manner. However, water extract of papaya did not affect cytotoxicity when compared with control. Water extract of papaya inhibited the phosphorylation of p38 and JNK induced by RANKL. The mRNA expression of c-Fos, NFATcl, TRAP and OSCAR induced by RANKL was inhibited by water extract of papaya treatment. Also, water extract of papaya suppressed the protein expression of c-Fos and NFATc1 in BMMs treated with RANKL. Taken together, these results suggest that papaya may be a useful drug in the treatment of bone-related disease.

Effect of Jak2 Inhibitor AG490 on Osteoclast Differentiation / 대한해부학회지

Osteoclasts are cells of hemopoietic origin that play an critical role in bone resorption and responsible for bone diseases, including osteoporosis, periodontal disease, and rheumatoid arthritis. In this study, we examined the effect of AG490, a Jak2-specific inhibitor on osteoclast differentiation. AG490 significantly inhibited receptor activator of NF-kappaB ligand (RANKL)-mediated osteoclast differentiation in a dose-dependent manner. RANKL stimulated the phosphorylation of p38, ERK, and JNK and promoted I-kappaB degradation. However, AG490 suppressed the phosphorylation of p38 induced by RANKL treatment. AG490 suppressed the mRNA expression of TRAP, c-Fos, NFATc1, and OSCAR in bone marrow-derived macrophages (BMMs) treated with RANKL. Also, AG490 significantly inhibited the protein expression of c-Fos and NFATc1 in response to RANKL. These results suggest that AG490 inhibited osteoclast differentiation by suppressing the expression of c-Fos and NFATc1.

CoPPIX Protects against TNBS Induced Colitis Through HO-1 Induction / 대한해부학회지

Crohn`s disease is a severe chronic inflammation that is treated mainly by immunosuppression, which often has serious side effects. There is a need to develop new drugs for treating this disease that have few side effects. Heme oxygenase-1 (HO-1) has immunosuppressive properties, but the mechanism of its anti-inflammatory actions is unclear. We investigated the protective effects of HO-1 on trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. An HO-1 inducer, cobalt protoporphyrin IX (CoPPIX), dramatically improved the clinical and histopathological symptoms in TNBS-induced colitis. CoPPIX suppressed tumor necrosis factor-alpha and interleukin-1beta expression and down-regulated the nuclear transcription factor kappa B activity caused by TNBS. The vehicle copper protoporphyrin IX (CuPPIX) failed to duplicate the protective effects seen with CoPPIX. Moreover, an inhibitor of HO-1 activity-zinc protoporphyrin IX-reversed the protective effects of CoPPIX on TNBS-induced colitis. In conclusion CoPPIX protects against TNBS-induced colonic damage by inducing HO-1, which might be an important target in the treatment of Crohn`s disease.