• Title/Summary/Keyword: phosphatase activity

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Effect of Lactobacillus acidophilus NS1 on the Hepatic Glycogen Contents in High-Fat Diet-Fed Mice (고지방식이 마우스의 간에서 Lactobacillus acidophilus NS1에 의한 글리코겐 함량 조절 효과)

  • Yang, Garam;Kim, Soyoung;Kim, Eungseok
    • Journal of Dairy Science and Biotechnology
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    • v.39 no.2
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    • pp.78-85
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    • 2021
  • Previously, we showed that oral administration of probiotics, Lactobacillus acidophilus NS1 (LNS1), improved insulin sensitivity in high-fat-diet-fed mice (HFD mice). Furthermore, LNS1-conditioned media (LNS1-CM) reduced HNF4α transcription activity and the expression of phosphoenol pyruvate carboxykinase (PEPCK), a key enzyme in gluconeogenesis in HepG2 cells. In this study, we demonstrated that LNS1 administration increased the expression of glycosyltransferase 2 (GYS2) and glucose transporter 2 (GLUT2), while reduced the expression of glucose-6-phosphatase (G6PC) expression in liver of HFD mice. Furthermore, LNS1 suppressed hepatic expression of glucokinase regulatory unit (GCKR) in HFD mice without changing the mRNA levels of glucokinase (GCK), suggesting that LNS1 may inhibit nuclear GCK activity. Consistently, addition of LNS1-CM to HepG2 cells increased the mRNA levels of GYS2 and GLUT2 with reduced mRNA levels of G6PC and GCKR. Moreover, hepatic glycogen contents were increased in HFD mice upon administration of LNS1. Together, these results suggest that LNS1 facilitates glycogen accumulation in liver by regulating the expression of genes involved in glycogen metabolism, contributing to improved insulin sensitivity in the HFD mice.

EFFECT OF ENAMEL MATRIX DERIVATIVE (EMD, $EMDOGAIN^{(R)}$) ON THE DIFFERENTIATION OF CULTURED HUMAN PERIODONTAL LIGAMENT CELLS AND MESENCHYMAL STEM CELLS (배양된 사람 치주인대세포와 골수유래간엽줄기세포의 분화에 미치는 법랑기질유도체 (Enamel Matrix Derivative, EMD)의 영향)

  • Park, Sang-Gyu;Jue, Seong-Suk;Kwon, Yong-Dae;Choi, Byung-Joon;Kim, Young-Ran;Lee, Baek-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.4
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    • pp.281-286
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    • 2009
  • Introduction: Enamel matrix derivative (EMD) is a protein which is secreted by Hertwig root sheath and plays a major role in the formation of cementum and attachment of peridontium. Several studies have shown that EMD promoted the proliferation and differentiation of preosteoblasts, osteoblasts and periodontal ligament cells in vitro: however, reports showing the inhibition of osteogenic differentiation by EMD also existed. This study was designed to simultaneously evaluate the effect of EMD on the two cell lines (human mesenchymal stem cells: hMSC, human periodontal ligament derived fibroblasts: hPDLCs) by means of quantitative analysis of some bone related matrices (Alkaline phosphatase : ALP, osteopontin ; OPN, osteocalcin ; OC). Materials and Methods: hMSCs and hPDLCs were expanded and cells in the 4${\sim}$6 passages were adopted to use. hMSc and hPDLCs were cultured during 1,2,7, and 14 days with 0, 50 and 100 ${\mu}g/ml$ of EMD, respectively. ALP activity was assessed by SensoLyte ALP kit and expressed as values of the relative optical density. Among the matrix proteins of the bony tissue, OC and OPN were assessed and quantification of these proteins was evaluated by means of human OC immunoassay kit and human OPN assay kit, respectively. Results: ALP activity maintained without EMD at $1,2^{nd}$ day. The activity increased at $7^{th}$ day but decreased at $14^{th}$ day. EMD increased the activity at $14^{th}$ day in the hPDLCs culture. In the hMSCs, rapid decrease was noted in $7^{th}$ and $14^{th}$ days without regard to EMD concentrations. Regarding the OPN synthesis in hPDLCs, marked decrease of OPN was noted after EMD application. Gradual decrease tendency of OPN was shown over time. In hMSCs, marked decrease of OPN was also noted after EMD application. Overall concentration of OPN was relatively consistent over time than that in hPDLCs. Regarding the OC synthesis, in both of hPDLCs and hMSCs, inhibition of OC formation was noted after EMD application in the early stages but EMD exerted minimal effect at the later stages. Conclusion: In this experimental condition, EMD seemed to play an inhibitory role during the differentiation of hMSCs and hPDLCs in the context of OC and OPN formation. In the periodontium, there are many kinds of cells contributing to the regeneration of oral tissue. EMD enhanced ALP activity in hPDLCs rather than in hMSCs and this may imply that EMD has a positive effect on the differentiation of cementoblasts compared with the effect on hMSCs. The result of our research was consistent with recent studies in which the authors showed the inhibitory effect of EMD in terms of the differentiation of mineral colony forming cells in vitro. This in vitro study may not stand for all the charateristics of EMD; thus, further studies involving many other bone matrices and cellular attachment will be necessary.

Effects of Compounds Isolated from an Ethanol Extract of the Sclerotium of Wolfiporia hoelen on Osteoblast Differentiation and Osteoclast Formation (복령 균핵의 에탄올 추출물에서 분리한 화합물의 조골세포 분화 촉진 및 파골세포 생성 억제 효과)

  • Sora Lee;Seokju Kim;Bowook Moon;Sik-Won Choi;Rhim Ryoo;Hyung Won Lee
    • Journal of Korean Society of Forest Science
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    • v.113 no.1
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    • pp.73-87
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    • 2024
  • Wolfiporia hoelen (Fr.) Y.C.Dai & V. Papp, commonly known as Poria cocos, is a significant traditional herb used for medicinal and culinary purposes Asian and European countries. Many studies have confirmed that the main components of W. hoelen have pharmacological activities and thatits extract has been shown to affect bone metabolism. This study aimed to the potential of a 50% ethanol extract of the sclerotium of W. hoelen for preventing and treating bone diseases. The ethanol extract was systematically fractionated using n-hexane, dichloromethane, and ethyl acetate. The dichloromethane fraction caused an approximately 29% increase in alkaline phosphatase (ALP) differentiation activity in C2C12 cells compared to the control. Four compounds isolated from this active dichloromethane fraction were identified through instrumental analysis and literature references as 3α-dehydrotrametenolic acid, ergosterol, pachymic acid, and dehydrotumulosic acid. All four compounds were evaluated at increasing concentrations (1, 3, 10, 30, and 100 μM) to determine their effects on ALP differentiation activity in C2C12 cells and RANKL-induced inhibition activity in bone marrow macrophages (BMMs), with a concurrent assessment of cytotoxicity at these concentrations. At a concentration of 3 μM, dehydrotumulosic acid caused a 160% increase in ALP activity, 24% higher than in the BMP-2 control. BMMs treated with dehydrotumulosic acid at concentrations between 10 and 100 μM showed a substantial 15-86% decrease in RANKL-induced inhibition activity compared to the control, with distinct patterns of RANKL inhibition and cytotoxicity observed at 10 μM. These findings suggest that the ethanol extract from the sclerotium of W. hoelen has potential to modulate bone-cell differentiation, while highlighting the possible benefits of dehydrotumulosic acid isolated from the dichloromethane fraction of W. hoelen for preventing and treating osteoporosis.

The Effects of Prostaglandin and Dibutyryl cAMP on Osteoblastic Cell Activity and Osteoclast Generation (Prostaglandin과 Dibutyryl cAMP가 조골세포의 활성과 파골세포 형성에 미치는 영향)

  • Mok, Sung-Kyu;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.448-468
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    • 1996
  • To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

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Effect of Pine (Pinus densiflora) Needle Extracts on Antioxidant Activity and Proliferation of Osteoclastic RAW 264.7 Cells (적송잎 추출물이 항산화 활성 및 파골세포의 증식에 미치는 영향)

  • Jeon, Min-Hee;Park, Mi-Ra;Park, Yong-Soo;Hwang, Hyun-Jung;Kim, Sung-Gu;Lee, Sang-Hyeon;Kim, Mi-Hyang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.4
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    • pp.525-530
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    • 2011
  • Pine needles have long been used as a traditional health-promoting medicinal food in Korea. This study was carried out to investigate the effects of pine needle extracts on the antioxidant activity, and proliferation of osteoclastic RAW 264.7 cells. Pine needle extracts were examined using hot water, ethanol, hexane, hot water-ethanol, and hot water-hexane. The effects of the pine needle extracts were examined by comparing the results with that of a commercial agents, proanthocyanidin. Analysis of each extract indicated that hot water-ethanol and ethanol extracts contained the highest total polyphenol concentrations. The hot water-ethanol and ethanol extracts also showed relatively the highest SOD-like activity. The proliferation of osteoclastic RAW 264.7 cells treated with pine needle extracts was decreased by lower than 70%. In addition, the hot water and ethanol extracts of pine needle significantly reduced the number of tartrate-resistant acid phosphatase-positive ($TRAP^+$) multinucleated cells from osteoclatic RAW 264.7 cells. These results indicate that pine needle extracts had an anabolic effect on bone through the promotion of osteoclast differentiation, suggesting that they could be used for the treatment of common metabolic bone diseases.

Induction of G1 Arrest by Methanol Extract of Lycopus lucidus in Human Lung Adenocarcinoma A549 Cells (택란 메탄올 추출물에 의한 인체 폐암 세포주 A549의 G1 arrest 유발)

  • Park, Hyun-Jin;Jin, Soojung;Oh, You Na;Yun, Seung-Geun;Lee, Ji-Young;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1109-1117
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    • 2013
  • Induction of G1 Arrest by Methanol Extract of Lycopus lucidus in Human Lung Adenocarcinoma A549 Cells Lycopus lucidus, a herbaceous perennial, is used as a traditional remedy in East Asia, including China and Korea. It has been reported that L. lucidus has anti-allergic effects, inhibitory effects on cholesterol acyltransferase in high glucose-induced vascular inflammation, and anti-proliferative effects in human breast cancer cells. However, the molecular mechanisms of the anti-cancer effects of L. lucidus have not yet been fully determined. In this study, we evaluated the anti-cancer effect and the mechanism of action of L. lucidus in human lung adenocarcinoma A549 cells using methanol extracts of L. lucidus (MELL). MELL treatment showed cytotoxic activity in a dose-dependent manner and induced G1 arrest in A549 cells. The induction of G1 arrest by MELL was associated with the up-regulation of phospho-CHK2 and the down-regulation of Cdc25A phosphatase. In addition, MELL treatment induced decreased expression of G1/S transition-related proteins, including CDK2, CDK4, CDK6, cyclin D1 and cyclin E. MELL also regulated the mRNA expression of CDK2 and cyclin E. On the other hand, the expression of p53 and the cyclin-dependent kinase inhibitor p21 was not induced by MELL. Collectively, these results suggest that MELL may exert an anti-cancer effect by cell cycle arrest at G1 phase through the ATM/CHK2/Cdc25A/CDK2 pathway in A549 cells.

Effects of Colpomenia sinuosa Extract on Serum Lipid Level and Bone Formation in Ovariectomized Rats (불레기말 추출물이 난소를 절제한 갱년기 장애 모델의 혈청 지질 변화 및 골 형성에 미치는 영향)

  • Lee, Juyeong;Kim, Bokyung;Park, Mi-Hwa;Choi, Kyung-Ha;Kong, Changsuk;Lee, Sang-Hyeon;Kim, Yuck Yong;Yu, Ki Hwan;Kim, Mihyang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.492-500
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    • 2016
  • In this study, the effects of Colpomenia sinuosa (CS) extract on serum lipid level and bone formation in an ovariectomized animal model were investigated. Twenty four nine-week old female Sprague-Dawley rats were randomly assigned to four groups: sham-operated rats (SHAM), ovariectomized control rats (OVX-CON), and ovariectomized rats supplemented with CS extract at 50 mg/kg bw (OVX-CS50) or 200 mg/kg bw (OVX-CS200). Three OVX groups were surgically ovariectomized while the SHAM group was sham-operated. CS extract was orally administrated at 1 mL per day. Analysis of serum lipid contents found that the total cholesterol, triglyceride, and low density lipoprotein cholesterol levels in the OVX-CON group were higher than those in the SHAM group. Notably, upon administration of CS extract after ovariectomy, triglyceride levels tended to significantly decrease. In addition, platelet aggregation ability decreased in groups treated with CS extract compared to the OVX-CON group. Serum alkaline phosphatase activity as an indicator of bone formation was lower in the CS extract group compared to the OVX-CON group. Collagen contents in bone and cartilage were reduced by ovariectomy, whereas the CS extract-supplemented groups exhibited higher concentrations in bone. According to these results, CS extract improved serum lipid parameters and osteogenesis in ovariectomized rats.

The Effect of Eisenia bicyclis Extracts on Bone Tissues in Ovariectomized Rats (대황 추출물이 갱년기 유도 흰쥐의 골 조직에 미치는 영향)

  • Park, Yong Soo;Kang, Min Suk;Kim, Bo Kyung;Kim, Mihyang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.1
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    • pp.33-39
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    • 2013
  • Menopause is often associated with several chronic diseases, including osteoporosis, cardiovascular disease, and obesity. In this study, we investigated the ability of Eisenia bicyclis (EB) to prevent bone loss in ovariectomized rats, a model for postmenopausal osteoporosis. Extracts from EB obtained using ethanol or hot water were analyzed for total polyphenol content and osteoporosis effects in vivo. Total polyphenol content was higher with extraction by hot water compared to ethanol extraction. Fifty 8-week-old female Sprague-Dawley rats were randomly assigned to four groups: the group were sham-operated rats (SHAM), ovariectomized rats (OVX-CON), and ovariectomized rats that were treated with EB at 50 mg/kg body weight (OVX-EB50) and 200 mg/kg body weight (OVX-EB200), respectively. The diets were fed to rats for 6 weeks after their operation. We found that the alkaline phosphatase (ALP) activity was lower in the EB extract group compared to the OVX-CON group. Collagen and pyridinoline content, in bone and cartilage, were reduced by ovariectomy, but the supplemented EB extract groups exhibited higher concentrations in their bones. These results suggest that EB can be used for the industrial development of foods with therapeutic functions.

Effect of Ecklonia stolonifera Extracts on Bone Turnover Markers in Ovariectomized Rats (곰피가 갱년기 장애 유도 흰쥐의 골 대사 지표물질의 변화에 미치는 영향)

  • Kim, Young-Kyoung;Kim, Mi-Hyang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1769-1775
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    • 2010
  • Menopause is often associated with the incidence of several chronic diseases including osteoporosis, cardiovascular disease, and obesity. The purpose of this study was designed to evaluate the possibility of osteoporosis prevention in postmenopausal women. In this study, we investigated the effects of Ecklonia stolonifera (ES) extracts on bone turnover markers in ovariectomized rats. For this study, the following four groups of 9-week-old Sprague-Dawley rats were evaluated over 6 weeks: normal rats (SHAM), ovariectomized rats (OVX-CON) and ovariectomized rats that were treated with ES extracts. We measured the osteocalcin and C-telopeptide of collagen cross-links (CTx) content, enzyme ALP activity in serum and collagen content in the cartilage, bone, skin and lungs. We found that the levels of indicators of bone metabolism such as alkaline phosphatase (ALP), osteocalcin and CTx were lower in rats in the ES extract group than the OVX-CON group. In addition, the collagen contents in the bone, cartilage, skin and lungs decreased in response to ovariectomy, but the levels of collagen were greater in the bone of rats that were treated with ES extract than in the bone of rats in the OVX-CON group. These results suggest that the ES may be an effective functional food to prevent osteoporosis in postmenopausal women.

Effect of exercise and energy restriction on blood pattern in obese children (운동과 에너지 제한이 비만아동의 혈액 성상에 미치는 영향)

  • 이성숙;오승호
    • Korean Journal of Human Ecology
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    • v.4 no.1
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    • pp.68-78
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    • 2001
  • The purpose of this study was to examine the effect of exercise and diet control program on blood pattern in obese children. The subjects of this study were five obese boys aged from 8 to 12 participating in the control period(C) for 3 days, exercise period(E) for one week, and exercise+energy restriction period(EER) for another one week, consecutively. The intensity of the exercise was 60~70% of HRmax and the energy was restricted at 493kcal/day. Daily mean total energy intake was 2,152${\pm}$138kcal. 1,861${\pm}$138kca1, and 1,368${\pm}$87kcal for the period C, E and EER, respectively. Body weight of after the program was significantly decreased from 48.94$\pm$5.11kg to 45.94${\pm}$4.74kg(P<0.01). And skinfold thickness. %fat. lean body mass, body mass index were significantly decreased(p<0.01). Blood sugar concentration was not significantly affected by weight loss, but alkaline phosphatase activity was significantly decreased. Concentrations of total lipid, LDL-cholesterol, triglyceride, phospholipid were not significantly decreased. But concentrations of HDL-cholesterol, %HDL-cholesterol, free fatty acid were significantly increased and concentration of VLDL-cholesterol, atherogenic index were significantly decreased. The results of this study showed that the obese children had a tendency to decrease coronary heart disease risk in the respect of plasma HDL-cholesterol and VLDL-cholesterol concentration by the exercise plus energy restriction program. Thus if we apply the lower intensity or duration of exercise for them this program might be more effective on the obese children.

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