• Title/Summary/Keyword: AMP-activated protein kinase

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THE EFFECT OF SODIUM FLUORIDE ON THE PHYSIOLOGICAL ROLE OF OSTEOBLASTIC CELL (불화나트륨이 조골세포의 생리적 활성에 미치는 영향)

  • Kim, Dae-Eop
    • Journal of the korean academy of Pediatric Dentistry
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    • v.25 no.3
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    • pp.635-648
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    • 1998
  • The clinical use of fluoride with a well known osteogenic action in osteoporotic patients is rational, because this condition is characterized by impaired bone formation. However, its anabolic effect has not been demonstrated well in vitro. The purpose of this study was to investigate the effects of sodium fluoride on the physiological role of osteoblastic cell. Osteoblastic cells were isolated from fetal rat calvaria. The results were as follows : 1. Mineralized nodules were shown in osteoblastic cell cultures, which had been maintained in the presence of ascorbic acid and ${\beta}-glycerophosphate$ up to 21 days. When cultures were treated with pulses of 48 hr duration before apparent mineralization was occurring, 2-fold increased in their number was detected. 2. Alkaline phosphatase activity of osteoblastic cells was inhibited by sodium fluoride in dose dependent manner. 3. The effect of sodium fluoride on the osteoblastic cell proliferation was measured by the incorporation of $[^3H]$-thymidine into DNA. As a result, sodium fluoride at $1{\sim}100{\mu}M$ increased the $[^3H]$-thymidine incorporation into DNA in a dose dependent manner. 4. The signaling mechanism activated by sodium fluoride dose-dependently enhanced the tyrosine phosphorylation of the adaptor molecule $Shc^{p66}$ and their association with Grb2, one of earlier events in a MAP kinase activation pathway cascade used by a significant subset of G protein-coupled receptors. 5. The phosphorylation of CREB(cAMP response element binding protein)was inhibited by the sodium fluoride in MC3T3E1 cells. In conclusion, the results of this study suggested that the mitogenic effect of the sodium fluoride in MC3T3E1 cell was stimulated in a dose-dependent manner and suggested "an important role for the interaction between She and Grb2" in controlling the proliferation of osteoblasts.

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Protective effects against alcoholic liver damage: potential of herbal juice (HJ), blend of Zingiber officinale Roscoe and Pueraria lobata Ohwi extracts

  • Young Yun Jung;You Yeon Choi;Woong Mo Yang;Kwang Seok Ahn
    • Journal of Convergence Korean Medicine
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    • v.5 no.1
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    • pp.45-54
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    • 2023
  • Objectives : Alcohol-induced liver disease advances as to reactive oxygen species (ROS) and cellular lipid peroxidation increase. We examined the hepatoprotective effects of Zingiber officinale Roscoe rhizome extract (ZR), Pueraria lobata Ohwi flower extracts (PF), and a newly developed herbal juice (HJ), which was a combination of ZR and PF extracts, against ethanol-induced hepatotoxicity. Methods: The study utilized the human hepatoma cell line HepG2 cells to validate the hepatoprotective effect of HJ (50~200 ㎍/mL) against ethanol (EtOH, 700 mM)-induced liver damage. Results: HJ effectively reduced the protein expression of sterol regulatory element-binding transcription factor 1, adiponectin, and AMP-activated protein kinase in EtOH-induced HepG2 cells. The levels of ROS, total cholesterol, and triglycerides, which are the result of various synthesis and lipogenesis processes induced by EtOH in the liver, were reduced by HJ. Furthermore, the activities of alcohol dehydrogenase and aldehyde dehydrogenase, enzymes linked to alcohol degradation, were more effectively downregulated by HJ treatment compared to treatment with ZR and PF alone, all without causing cytotoxic effects. Conclusions: HJ protects the liver by inhibiting EtOH-induced lipogenesis, lowering ROS generation, and improving alcohol degradation, which is more effective than ZR and PF alone. Further, in vivo experiments can offer additional evidence regarding the effectiveness, safety, and underlying mechanism of action of HJ.

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Mountain-cultivated Ginseng Ripened into Persimmon Vinegar Ingestion on Fat Storage and Metabolic Protein Expression in Diet-controlled Rats (산양삼 혼입 숙성 감식초 섭취에 의한 식이 제한 흰쥐의 지방 저장 및 에너지 대사 단백질 발현)

  • Lee, In-Ho;Kim, Pan-Ki;Ryu, Sungpil
    • Journal of Korean Society of Forest Science
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    • v.104 no.1
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    • pp.67-75
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    • 2015
  • This research is to investigate the four years growth mountain-cultivated ginseng ripened twenty-two weeks into four years fermented persimmon vinegar (tentatively: Sansamcho) ingestion on obese-related factors during dietary control. The Sansamcho was ingested orally, two times a day, after every meal for six weeks to the male rats. Groups were divided into the control (CON), the restricted diet (RD), and the weight cycling (WC). And, each groups has its own sub-groups as the -control (-CON), 2.5 times diluted Sansamcho ingestion (-MPV2.5), and 5.0 times diluted Sansamcho ingestion (-MPV5.0) groups, respectively. The number of rat was consisted of seven in each group. After six weeks rearing periods was done, abdominal fats (retroperitoneal fat, mesentery fat, and epididymal fat) and energy metabolic-related protein (AMPK: AMP-activated protein kinase; PPAR-${\alpha}$: peroxisome proliferator-activated receptor-${\alpha}$; and CPT-1: carnitine palmitoyltransferase-1) were weighed and analyzed. Amount of stored fat was significantly or tended to decrease by Sansamcho ingestion. In addition, sum of fats increasing were suppressed by the material. On the contrary, energy metabolism-related protein expression was significantly increased or tended to increase by Sansamcho ingestion. This results suggested that increased energy metabolism using Sansamcho was restrained effectively visceral fat store by high-fat diet and/or dietary control. In other words, it has a good function to suppress weight cycling which is the most insoluble problem. Therefore, the fusion material, Sansamcho, may expect to utilize as the obese-suppression-food.

Mechanisms of Insulinotropic Effect of YHB-2017 [Genistein] Isolated from fermentation Broths of Streptomyces sp. (방선균에서 유래한 YHB-2017 [Genistein]의 인슐린 분비 촉진 작용 기전)

  • Kwag, Won-Jae;Park, You-Hoi;Park, Jun-Chul;Lee, Byung-Kyu;Kang, Yup;Choe, Tae-Boo
    • KSBB Journal
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    • v.21 no.6 s.101
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    • pp.466-473
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    • 2006
  • Impaired insulin secretion from pancreatic beta-cells in response to glucose is an important feature in the pathology of non-insulin-dependent diabetes mellitus (NIDDM). In the course of screening for useful insulin secretagogues, we have isolated and identified YHB-2017 (Genistein) as a insulin secretion potentiator from fermentation broths of our in-house microbial library. The insulinotropic activity of YHB-2017 in isolated rat pancreatic islets was exerted only at high concentration of glucose (8.3-16 mM) but not at low concentration of glucose (3.3-5.5 mM). Also, in perifusion study with isolated rat pancreatic islets, YHB-2017 stimulated insulin secretion in a time-dependent manner when YHB-2017 was added to KRB buffer containing 16 mM glucose. In the presence of $200\;{\mu}M$ diazoxide and 35 mM KCI, which stimulates maximum $Ca^{2+}$ influx independently of KATP channel, YHB-2017 enhanced KATP channel-independent insulin secretion at high concentration glucose (16 mM). To elucidate the mechanisms of the glucose-dependent potentiation effect of YHB-2017, pharmacologic inhibitors for protein kinase A, protein kinase C and calcium/calmodulin kinase II were pre-treated and then the potentiation effect of YHB-2017 on insulin secretion was investigated. Pre-treatment of H89 as a PKA inhibitor had a significant inhibitory effect on YHB-2017-induced potentiation effect. Furthermore, western immunoblotting analyses revealed that YHB-2017 increased phosphorylation of PKA substrates and cAMP response element-binding protein (CREB) under high concentration of glucose. These results demonstrated that the insulinotropic effect of YHB-2017 is mediated through PKA signal pathway and activated amplifying $K_{ATP}$ channel-independent insulin secretion pathway.

Effects of N-acetylcysteine on the energy status and antioxidant capacity in heart and liver of cold-stressed broilers

  • Li, Chengcheng;Peng, Meng;Liao, Man;Guo, Shuangshuang;Hou, Yongqing;Ding, Binying;Wu, Tao;Yi, Dan
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.9
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    • pp.1444-1454
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    • 2020
  • Objective: Cold stress induces oxidative damage and impairs energy status of broilers. N-acetylcysteine (NAC) exhibits antioxidant properties and modulates energy metabolism of animals. This study was conducted to investigate the effects of NAC on energy status and antioxidant capacity of heart and liver in the cold-stressed broilers. Methods: The experiment consisted of 4 treatments in a 2×2 factorial arrangement with two diets (basal diet or plus 0.1% NAC) and two ambient temperatures (thermoneutral [conventional ambient temperature] or cold stress [10℃±1℃ during days 15 to 42]). Results: No ascites were seen in cold-stressed broilers. NAC did not attenuate the impaired growth performance of stressed birds. However, NAC decreased plasma asparagine but increased aspartate levels in cold-stressed birds (p<0.05). NAC reduced hepatic adenosine triphosphate (ATP) but elevated adenosine diphosphate contents in unstressed birds (p<0.05). The hepatic ratio of adenosine monophosphate (AMP) to ATP was increased in birds fed NAC (p<0.05). NAC decreased plasma malondialdehyde (MDA) level and cardiac total superoxide dismutase (T-SOD) activity in unstressed birds, but increased hepatic activities of T-SOD, catalase and glutathione peroxidase in stressed birds (p<0.05). NAC down-regulated hepatic AMP-activated protein kinase but up-regulated cardiac heme-oxigenase mRNA expression in stressed birds, and decreased expression of hepatic peroxisome proliferator-activated receptor coactivator-1α as well as hypoxia-inducible factor-1α in liver and heart of birds. Conclusion: Dietary NAC did not affect energy status but enhanced the hepatic antioxidant capacity by increasing the activities of antioxidant enzymes in cold-stressed broilers.

The Expression of Adipogenic Genes in Adipose Tissues of Feedlot Steers Fed Supplementary Palm Oil or Soybean Oil

  • Choi, Seong Ho;Park, Sung Kwon;Choi, Chang Weon;Li, Xiang Zi;Kim, Kyoung Hoon;Kim, Won Young;Jeong, Joon;Johnson, Bradley J.;Zan, Linsen;Smith, Stephen B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.404-412
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    • 2016
  • We hypothesized that supplementing finishing diets with palm oil would promote adipogenic gene expression and stearoyl-CoA desaturase (SCD) gene expression in subcutaneous (s.c.) and intramuscular (i.m.) adipose tissues of feedlot steers. Eighteen Angus and Angus crossbred steers were assigned to three groups of 6 steers and fed a basal diet (control), with 3% palm oil, or with 3% soybean oil, for 70 d, top-dressed daily. Tailhead s.c. adipose tissue was obtained by biopsy at 14 d before the initiation of dietary treatments and at 35 d of dietary treatments. At slaughter, after 70 d of dietary treatment, tailhead s.c. adipose tissue and i.m. adipose tissue were obtained from the longissimus thoracis muscle. Palm oil increased plasma palmitic acid and soybean oil increased plasma linoleic acid and ${\alpha}$-linolenic acid relative to the initial sampling time. Expression of AMP-activated protein kinase alpha ($AMPK{\alpha}$) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) increased between the initial and intermediate biopsies and declined thereafter (p<0.03). SCD gene expression did not change between the initial and intermediate biopsies but declined by over 75% by the final period (p = 0.04), and G-coupled protein receptor 43 (GPR43) gene expression was unaffected by diet or time on trial. Soybean oil decreased (p = 0.01) $PPAR{\gamma}$ gene expression at the intermediate sample time. At the terminal sample time, $PPAR{\gamma}$ and SCD gene expression was less in i.m. adipose tissue than in s.c. adipose tissue (p<0.05). $AMPK{\alpha}$ gene expression was less in s.c. adipose tissue of palm oil-fed steers than in control steers (p = 0.04) and CCAAT enhancer binding protein-beta ($CEBP{\beta}$) gene expression was less in s.c. and i.m. adipose tissues of palm oil-fed steers than in soybean oil-fed steers (p<0.03). Soybean oil decreased SCD gene expression in s.c. adipose tissue (p = 0.05); SCD gene expression in palm oil-fed steers was intermediate between control and soybean oil-fed steers. Contrary to our original hypothesis, palm oil did not promote adipogenic gene expression in s.c. and i.m. adipose tissue.

Effects of Polygonatum sibiricum rhizome extract on lipid and energy metabolism in high-fat diet-induced obese mice (고지방 식이 유도 비만 마우스 모델에서 황정 추출물의 지방질 및 에너지 대사 관련 유전자에 대한 효능 연구)

  • Jeon, Woo-Jin;Kim, Ji-Young;Oh, Ik-Hoon;Lee, Do-Seop;Shon, Suh-Youn;Seo, Yun-Ji;Yeon, Seung-Woo;Kang, Jae-Hoon
    • Korean Journal of Food Science and Technology
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    • v.49 no.2
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    • pp.192-202
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    • 2017
  • In this study, factors involved in lipid and energy metabolism following treatment with ethanolic extract of the Polygonatum sibiricum rhizome (ID1216) were evaluated in high-fat diet-induced obese mice. ID1216-treated mice showed a significant reduction in weight gain compared to non-treated mice. ID1216 treatment increased the protein levels of AMP-dependent protein kinase, sirtuin1, peroxisome proliferator-activated receptor ${\gamma}$ coactivator 1-${\alpha}$ ($PGC1{\alpha}$), peroxisome proliferator-activated receptor ${\alpha}$ ($PPAR{\alpha}$) and uncoupling proteins in the adipose tissue, liver and muscle compared to vehicle treatment. Analysis of downstream signals of the sirtuin1 $PGC1{\alpha}$-$PPAR{\alpha}$ pathway showed that ID1216 regulates the expression of ${\beta}$-oxidation related genes such as acyl-CoA oxidase, carnitine palmitoyltransferase1, acyl-CoA dehydrogenase and adipocyte protein 2. In addition, ID1216 increased the expression of adipose triglyceride lipase. These results suggest that ID1216 has anti-obesity effects by regulating the genes involved thermogenesis, ${\beta}$-oxidation and lipolysis in a diet-induced obesity model.

Inhibition of Differentiation and Anti-Adipogenetic Effect of the Salvia plebeia R. Br. Ethanol Extract in Murine Adipocytes, 3T3-L1 Cells (배암차즈기 에탄올 추출물의 3T3-L1 지방전구세포 분화 억제 및 지방 축적 저해 효과)

  • Kim, Sung-Ok;Kim, Mi-Ryeo;Hwang, Kyung-A;Park, No-Jin;Jeong, Ji-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.4
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    • pp.401-408
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    • 2017
  • Salvia plebeia R. Br. (Lamiaceae) has been used in folk medicines in Asian countries, including Korea and China, to treat inflammatory diseases. The focus of our research was on the anti-adipogenic activity of ethanol extract from Salvia plebeia R. Br. (SPE) in 3T3-L1 adipocytes. This study investigated inhibition of differentiation and lipogenesis upon SPE treatment in 3T3-L1 cells. The results reveal that SPE at non-cytotoxic concentration significantly suppressed triglyceride accumulation and reduced expression of peroxisome proliferator-activated receptor gamma, CCAAT/enhancer-binding protein-alpha, and sterol regulatory element-binding protein as adipogenic transcription factors in 3T3-L1 adipocytes compared to non-treated control cells. Inducible phosphorylation of AMP-activated protein kinase, acetyl CoA carboxylase, and hormone-sensitive lipase as well as carnitine palmitoyltransferase-1 mRNA expression increased upon SPE treatment, which suppressed expression of fatty acid synthase. In conclusion, these results demonstrate that SPE can inhibit expression of adipogenic genes in 3T3-L1 adipocytes. Our study suggests that SPE has potential anti-obesity effects and is a novel therapeutic functional agent with anti-adipogenic activity via reduction of lipogenesis.

Red pepper seed water extract inhibits preadipocyte differentiation and induces mature adipocyte apoptosis in 3T3-L1 cells

  • Kim, Hwa-Jin;You, Mi-Kyoung;Lee, Young-Hyun;Kim, Hyun-Jung;Adhikari, Deepak;Kim, Hyeon-A
    • Nutrition Research and Practice
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    • v.12 no.6
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    • pp.494-502
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    • 2018
  • BACKGROUND/OBJECTIVES: Reducing the number of adipocytes by inducing apoptosis of mature adipocytes as well as suppressing differentiation of preadipocytes plays an important role in preventing obesity. This study examines the anti-adipogenic and pro-apoptotic effect of red pepper seed water extract (RPS) prepared at $4^{\circ}C$ (RPS4) in 3T3-L1 cells. MATERIALS/METHODS: Effect of RPS4 or its fractions on lipid accumulation was determined in 3T3-L1 cells using oil red O (ORO) staining. The expressions of AMP-activated protein kinase (AMPK) and adipogenic associated proteins [peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR-{\gamma}$), CCAAT/enhancer-binding proteins ${\alpha}$ (C/EBP ${\alpha}$), sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC)] were measured in 3T3-L1 cells treated with RPS4. Apoptosis and the expression of Akt and Bcl-2 family proteins [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated death promoter (Bad), Bcl-2 like protein 4 (Bax), Bal-2 homologous antagonist/killer (Bak)] were measured in mature 3T3-L1 cells treated with RPS4. RESULTS: Treatment of RPS4 ($0-75{\mu}g/mL$) or its fractions ($0-50{\mu}g/mL$) for 24 h did not have an apparent cytotoxicity on pre and mature 3T3-L1 cells. RPS4 significantly suppressed differentiation and cellular lipid accumulation by increasing the phosphorylation of AMPK and reducing the expression of $PPAR-{\gamma}$, C/EBP ${\alpha}$, SREBP-1c, FAS, and ACC. In addition, all fractions except ethyl acetate fraction significantly suppressed cellular lipid accumulation. RPS4 induced the apoptosis of mature adipocytes by hypophosphorylating Akt, increasing the expression of the pro-apoptotic proteins, Bak, Bax, and Bad, and reducing the expression of the anti-apoptotic proteins, Bcl-2 and p-Bad. CONCLUSIONS: These finding suggest that RPS4 can reduce the numbers as well as the size of adipocytes and might useful for preventing and treating obesity.

Effects of steamed Polygonatum odoratum extract on inhibition of adipocyte differentiation and lowing lipid in 3T3-L1 adipocytes (증자 둥굴레 추출물의 3T3-L1 지방세포에서 분화억제 및 지질강하 효과)

  • Kang, Byung Tae;Choe, Won Kyung;Park, Dong Cheol;Kim, Jong Kuk;Park, Mora;Kim, Sung Ok;Kim, Mi Ryeo
    • The Korea Journal of Herbology
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    • v.29 no.2
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    • pp.15-21
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    • 2014
  • Objectives : The purpose of this study was to investigate inhibitory effects of steamed Polygonatum odoratum extract (POE) on differentiation and adipogenesis in 3T3-L1 adipocytes. Methods : Polygonatum odoratum (P. odoratum) extract was extracted with ethyl acetate. Total phenolic and flavonoid contents in POE were measured for antioxidant activity. The spectrophotometric method was used to determine the DPPH and ABTS radical scavenging activity and ferric-reducing antioxidant potential (FRAP). MTT assay was examined for cell toxicity, oil red O staining was performed for intracelluar adipogenesis in differentiated 3T3-L1 adipocytes. Western blot analysis for measurement of CCAAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor${\gamma}$ ($PPAR{\gamma}$) and AMP-activated protein kinase (AMPK) expressions were performed. Results : The results revealed that POE has antioxidant activities. Contents of total polyphenolics and flavonoids were $50.83{\pm}1.52$ GAE mg/100g dry weight of POE and $17.05{\pm}2.47$ RE mg/100g dry weight of POE, respectively. DPPH radical scavenging activity, and FRAP in 10 mg/ml concentration were $92.1{\pm}0.6%$, $244.8{\pm}9.0{\mu}M$ Fe(II) and ABTS inhibition in 5 mg/ml concentration was $84.8{\pm}4.1%$. Treatment of POE in adipocytes inhibited the differentiation and adipogenesis of 3T3-L1 adipocytes compared to those of vehicle control. Additionally, protein expressions of $C/EBP{\alpha}$ and $PPAR{\gamma}$, major transcription factor for the adipogenic genes, were significantly decreased compared to those of vehicle control (p<0.05). Futhermore, phosphorylation of AMPK was increased in 3T3-L1 adipocytes treated with POE compared to that of vehicle control (p<0.05). Conclusions : we demonstrate that steamed P. odoratum extract (POE) has potentiating antioxidant activities, inhibits differentiation and lipid accumulation and also induces energy expenditure in adipocytes, which may contribute to antiobesity property.