• Title/Summary/Keyword: Lipid Binding

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Effects of a Herbal Preparation HJ01 on Adipocyte Differentiation in OP9 Cells and the Poloxamer-407 Induced Hyperlipidemia in Mice (HJ01이 OP9세포에서의 지방 분화와 P-407로 유발한 고지혈증 흰쥐에 미치는 영향)

  • Park, Jung-Eun;Han, Sang-Yong;Choi, Eun-Sik;Chong, Myong-Soo;Kim, Yun-Kyung
    • Herbal Formula Science
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    • v.21 no.1
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    • pp.99-110
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    • 2013
  • Objectives : This study was designed to investigate the effect of a herbal preparation HJ01 consisting of Salicornia herbacea, Citri Reticulatae Pericarpium, Crataegi Fructus and Glycyrrhizae Radix on adipocyte differentiation in OP9 cells and on poloxamer 407(P-407)-induced hyperlipidemia in mice. Methods : 1. MTT assay was used to evaluate the potential cytotoxicity of Salicornia herbacea, Citri Reticulatae Pericarpium, Crataegi Fructus, Glycyrrhizae Radix and HJ01, respectively. 2. Bone-marrow derived OP9 cells were treated with HJ01, and the alterations in fat storage in the cells were determined by the Oil red O assay. 3. The protein level of CAAAT/enhancer binding protein alpha($C/EBP{\alpha}$), as a adipocyte differentiation marker, was examined using western blot analysis in differentiated OP6 cells. 4. Adult male C57BL6 mice received intraperitoneal injections of P407 to induce hyperlipidemia, simultaneously, were treated with HJ01 for 4 weeks. Then the cholesterol (TC), triglyceride (TG) and high-density lipoprotein-cholesterol (HDL-c) levels in sera and liver tissues were measured. Results : 1. The MTT assay exhibited that Salicornia herbacea, Citri Reticulatae Pericarpium, Crataegi Fructus, Glycyrrhizae Radix and HJ01 showed no significant cytotoxicity in tested dosages. 2. Ten days' treatment with HJ01 markedly inhibited the increases in fat storage in differentiated OP6 cells. 3. Four weeks' treatment with HJ01 down-regulated the protein level of CAAAT/enhancer binding protein alpha($C/EBP{\alpha}$) but up-regulated the levels of adiponectin in differentiated OP9 cells. 5. HJ01 inhibited the accumulation of TC and TG in liver tissues and increased serum levels of TC in hyperlipidemic mice. Conclusions : These results suggest that HJ01 can in vitro inhibit adipocyte differentiation and fat storage in OP6 cells, in vivo improve the hyperlipidemia induced by P-407 in mice, which may be mediated by promoting glucose uptake and improving a lipid metabolite profile.

Prostaglandin E2 Reverses Curcumin-Induced Inhibition of Survival Signal Pathways in Human Colorectal Carcinoma (HCT-15) Cell Lines

  • Shehzad, Adeeb;Islam, Salman Ul;Lee, Jaetae;Lee, Young Sup
    • Molecules and Cells
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    • v.37 no.12
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    • pp.899-906
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    • 2014
  • Prostaglandin $E_2$ ($PGE_2$) promotes tumor-persistent inflammation, frequently resulting in cancer. Curcumin is a diphenolic turmeric that inhibits carcinogenesis and induces apoptosis. $PGE_2$ inhibits curcumin-induced apoptosis; however, the underlying inhibitory mechanisms in colon cancer cells remain unknown. The aim of the present study is to investigate the survival role of $PGE_2$ and whether addition of exogenous $PGE_2$ affects curcumininduced cell death. HCT-15 cells were treated with curcumin and $PGE_2$, and protein expression levels were investigated via Western blot. Reactive oxygen species (ROS) generation, lipid peroxidation, and intracellular glutathione (GSH) levels were confirmed using specific dyes. The nuclear factor-kappa B ($NF-{\kappa}B$) DNA-binding was measured by electrophoretic mobility shift assay (EMSA). $PGE_2$ inhibited curcumin-induced apoptosis by suppressing oxidative stress and degradation of PARP and lamin B. However, exposure of cells to the EP2 receptor antagonist, AH6809, and the PKA inhibitor, H89, before treatment with $PGE_2$ or curcumin abolished the protective effect of $PGE_2$ and enhanced curcumin-induced cell death. $PGE_2$ activates PKA, which is required for cAMP-mediated transcriptional activation of CREB. $PGE_2$ also activated the Ras/Raf/Erk pathway, and pretreatment with PD98059 abolished the protective effect of $PGE_2$. Furthermore, curcumin treatment greatly reduced phosphorylation of CREB, followed by a concomitant reduction of $NF-{\kappa}B$ (p50 and p65) subunit activation. $PGE_2$ markedly activated nuclear translocation of $NF-{\kappa}B$. EMSA confirmed the DNA-binding activities of $NF-{\kappa}B$ subunits. These results suggest that inhibition of curcumin-induced apoptosis by $PGE_2$ through activation of PKA, Ras, and $NF-{\kappa}B$ signaling pathways may provide a molecular basis for the reversal of curcumin-induced colon carcinoma cell death.

Study on Anti-obesity Effect of Chegameuiin-tang (체감의이인탕(體減薏苡仁湯)의 항비만 효과 연구)

  • Park, Tae-Yong;Shin, Byung-Cheul;Kong, Jae-Cheol;Song, Mi-Young;Kim, Eun-Kyung;Seo, Eun-A;Ryu, Do-Gon;Kwon, Kang-Beom
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.3
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    • pp.642-648
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    • 2008
  • The aim of this study is to investigate Chegameuiin-tang water extracts (CETE) have potent anti-obesity activities in a high fat diet-induced obesity mouse model. In this study, we designed three group (normal diet group, high fat diet group, high fat diet plus CETE group for 13-week oral administration). Increases in body weight and fat storage were inhibited by 13-week oral administration of CETE at a 500 mg/kg concentration in this animal model, while the amount of food intake was not affected. Results from blood lipid analysis showed that the levels of triglyceride, total cholesterol and LDL-cholesterol were significantly lowered by CETE administration, also HDL-cholesterol was increased more than high fat diet-induced obese mouse. To understand the underlying mechanism at the molecular level, the effects of CETE were examined on the expression of the genes involved in lipogenesis and lipolysis by real-time PCR. In epididymal fat of CETE-treated mice, the mRNA level of lipogenic genes such as sterol regulatory element binding protein 1 and fatty acid synthase were decreased, which was well correlated with the reduction of the epididymal fat weight. Also, CETE administration inhibited decreases of the hormone-sensitve lipase and lipoprotein lipase mRNA expressions, which are genes related with lipolysis. These results suggest that Chegameuiin-tang may have great potential as a novel anti-obesity agent.

Antioxidant Activity and Inhibitory Effect of Aster scaber Thunb. Extract on Adipocyte Differentiation in 3T3-L1 Cells (참취(Aster scaber Thunb.) 추출물의 항산화 효과와 3T3-L1 지방전구세포에서의 지방분화 억제 효과)

  • Choi, Jun-Hyeok;Park, Yun-Hee;Lee, In-Seon;Lee, Sam-Pin;Yu, Mi-Hee
    • Korean Journal of Food Science and Technology
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    • v.45 no.3
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    • pp.356-363
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    • 2013
  • Clinical and preclinical trials of involving drugs with anti-obesity effects have focused on screening for herbal medicines suspected to have anti-obesity activities. In this study, an extract of Aster scaver Thunb., which was prepared in 80% methanol (ASE), was assessed for its total phenol content, total flavonoid content, antioxidant activity ability to scavenge the ${\alpha}-{\alpha}$-diphenyl-${\beta}$-picrylhydrazyl, 2,2'-azino-bis-[3-ethylbenzthiazoline]-6-sulfonic acid radical, and anti-adipogenic effects. The anti-adipogenic effect of ASE on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated by assaying the suppression of adipocyte differentiation and lipid accumulation by using western blot analysis and the Oil Red-O assay, respectively. The staining results showed that ASE significantly inhibited 3T3-L1. Western blot analysis results showed that ASE decreased the levels of peroxisome proliferator-activated receptor-${\gamma}$, CCAAT/enhancer-binding protein ${\alpha}$, and sterol regulatory element-binding protein 1c. These results demonstrate that ASE directly inhibits the differentiation of preadipocytes, and might be an important adjunct in the therapeutic efforts to reduce adipogenesis.

Extracts of Housefly Maggot Reduces Blood Cholesterol in Hypercholesterolemic Rats (고콜레스테롤 랫드에서 파리유충 추출물의 혈액지질 감소기전)

  • Park, Byung-Sung;Park, Sang-Oh
    • Journal of the Korean Applied Science and Technology
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    • v.31 no.1
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    • pp.101-112
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    • 2014
  • The aim of this study was to evaluate the biological mechanism of orally administered ethanolic extract of fly maggot(EM) on hypocholesterolemic rats fed a high-cholesterol diet. Sprague Dawley male rats were divided into four groups (EM dose control=0, 5.0, 7.0, and 9.0 mg/100 g BW) and were treated for 6 weeks. EM groups revealed a significant reduction in serum triglyceride, total cholesterol, and LDL-C when compared with the control group(p<0.05). HMG-CoA reductase activity in EM groups were lower than those of the control group, but total sterol, neutral sterol, and bile acid excretion were increased in EM groups when compared with the control group(p<0.05). To identify the biological mechanism of EM towards the hypocholesterolemic effect, sterol response element binding proteins (SREBPs) and the peroxisome proliferator-activated receptors ($PPAR{\alpha}$ transcription system were determined in rats fed a high-cholesterol diet. It was discovered that EM suppress the expression of SREBP-$1{\alpha}$ and SREBP-2 mRNA in the liver tissues of high-cholesterol diet fed rats, while simultaneously increasing the expression of $PPAR{\alpha}$ mRNA(p<0.05). This finding indicates that EM may have hypocholesterolemic effects in rats fed a high-cholesterol diet, by regulating cholesterol metabolism-related biochemical parameters and SREBP-$1{\alpha}$ SREPB-2 and $PPAR{\alpha}$gene expression.

The Effects of Regulatory Exercise on Adipokines and Inflammatory Reaction in Type 1 Diabetic and Obese Children (규칙적인 운동이 제1형 당뇨병 및 비만 환아의 아디포카인과 염증반응에 미치는 영향)

  • Kang, Sung-Hwun;Shin, Ki-Ok;Park, So-Young;Woo, Jin-Hee
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1066-1072
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    • 2010
  • The purpose of this study was to investigate the effects of aerobic exercise on adipokines and inflammatory reaction in obesity and type 1 diabetes mellitus (T1DM) children. We studied obese (OG, n=9), type 1 diabetic (DG, n=9), and normal (NG, n=9) children groups. Measurement factors included body weight, % fat, body mass index (BMI), $VO_2max$, lipid profiles and adipokines. The results showed significant differences in body weight, % fat, BMI, and $VO_2max$ (ml/kg/min) among the OG, DG, and NG (p<0.05) groups. There were significant differences in LDL-C and HDL-C between the OG, DG, and NG groups (p<0.05). In addition, adiponectin and retinol binding protein (RBP)-4 were significantly changed in DG and NG after 12 weeks exercise training (p<0.05), and there were also significant differences among the OG, DG, and NG groups (p<0.05). Monocyte chemoattractant protein (MCP)-1 in the OG, DG, and NG groups was significantly increased after 12 weeks exercise training (p<0.05). In conclusion, regulatory aerobic exercise does not change body composition in obese children with T1DM, but exercise and decreased blood inflammatory factors in T1DM may protect obese children from metabolic syndrome.

Effect of Dietary Fibers on Retinol Binding Protein (RBP, cRBP I, cRBP II) Gene Expression in Rats Fed High Fat Diet (고지방 식이내 식이섬유질이 흰쥐의 레티놀 결합 단백질 (RBP, cRBP I, cRBP II) 유전자 발현에 미치는 영향)

  • Jo, Min-Wha;Shin, Dong-Soon
    • Journal of Nutrition and Health
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    • v.38 no.10
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    • pp.817-826
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    • 2005
  • Recently it has been reported that vitamin A and retinol binding proteins (RBPs) in blood and urine were changed in the condition of diabetes mellitus or hyperlipidemia. Fruits and vegetables are recommended to consume for the people suffered from these chronic degenerative diseases. The main components of fruits and vegetables are dietary fibers, for example cellulose and pectin, of which function to affect the absorption and excretion of dietary fat and fat-soluble substances. This study was conducted to investigate the effect of dietary fibers on RBPs mRNA expression in liver, small intestine and serum of rat fed high fat diet during 4 weeks. Sprague-Dawley rats, weighing 121g on average, were divided into four groups; (Control; $17\%$ fat & cellulose supplement diet, HF0: $25\%$ fat & fiber free diet, B:.Uc: $25\%$ fat & cellulose supplement diet and HF0: $25\%$ fat & pectin supplement diet) . The rats fed high fat diet groups (HF0, HFC, HFP) tended to consume the food less than the control group, but FER of HF0 groups was significantly higher than the control (p < 0.05) . The weight of adrenal gland in high fat diet groups (HF0, HFC, HFP) was significantly less than the control. Total lipid in feces daily excreted and in liver did not show any significant differences among the groups. Total cholesterol in HFP group was significantly different from that of HFC group. Serum total cholesterol and triglyceride in other group tended to lower than other groups and HDL cholesterol higher. Consequently, AI (atherogenic index) was the lowest in HFP group. Vit A contents in feces daily excreted tended to lower in high fat diet groups (HF0, HFP) compared to the control group. That content in adrenal gland was the lowest in HF0 group, but not in liver. In HFP group were down-regulated cRBPI mRNA in liver and cRBPII mRNA in small intestine and up-regulated RBP and transthyretin expression in serum compared to the other groups. In conclusion, dietary fibers, especially pectin, in high fat diet might down-regulate the expression of CRBP I, CRBP II mRNA in liver and small intestine, but increase the secretion of RBP into serum and therefore inhance the bioavailability of Vit A through the body. (Korean J Nutrition 38(10): 817$\sim$826,2005)

THE AFFINITY OF CALMODULIN-AFFIGEL FOR INOSITOL TRIPHOSPHATE KINASE FROM BOVINE BRAIN (소의 뇌 Inositol triphosphate kinase와 Calmodulin-Affigel과의 친화도)

  • Lim, Sung-Woo;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.7 no.1
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    • pp.39-50
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    • 1990
  • The one event on signalling mechanism is the cleavage by adenyl cyclase of ATP into second messenger, cyclic AMP. The other transfer system of inositol metabolism. it is widely recognized that hydrolysis of the minor membrane lipid phosphoinositide bisphosphate($PIP_2$) initiated by occupation of certain receptors and catalyzed by phospholipase C, lead to toe generation of the two intracellular messengers, inositol triphosphate($IP_3$) and diacylglycerol(DG). $IP_3$ is converted to inositol tetrakisphosphate($IP_4$) by $IP_3$ kinase. In the present study, it is that purification of calmodulin is used by phenyl-Sepharose CL-4B chromatography. it's molecular weigh, 17.000 in SDS-polyacrylamide gel electrophoresis. In order to observe the affinity between calmodulin (CaM)-Affigel 15 and $IP_3$ kinase, and isolated $IP_3$ kinase, was applied in CaM-Affigel with $Ca^{2+}$ equilibirum buffer and EGTA equilibirum buffer. We compared with binding and elution effect of $IP_3$ kinase in several condition of buffer. In affinity of binding. $Ca^{2+}$ equilibrium buffer was in the most proper condition. and elution, CaM/$Ca^{2+}$ buffer(CE1 10.36, CE2 12. 76pM/min/mg of protein) was effected much more than EGTA buffer(E2 1.48, E3 2.43pM/min/mg of protein), but CaM/$Ca^{2+}$ stimulate the activity of $IP_3$ kinase. And then, several detergents such as sodium deoxycholate, tween 20. cholic acid, polyethylene glycol, chaps were applied. The 0.2% chaps buffer(E2 23.19, E3 8.05pM/min/mg of protein) was the most effective in elution of $IP_3$ kinase.

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Inhibitory Effect of the Ethanol Extract of Torilis Japonica Decandolle on Adipocyte Differentiation in 3T3-L1 Cells (사상자 에탄올 추출물의 지방세포 분화 억제 효과)

  • Nam, Gun He;Wee, Ji-Hyang;Kim, Sang Yung;Baek, Ji-Young;Kim, Young Min
    • Journal of Life Science
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    • v.29 no.9
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    • pp.1016-1022
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    • 2019
  • Owing to increased interest in preventing obesity in an aging society, both men and women spend considerable amount of cost on obesity managements. In this study, we investigated the natural substances on anti-obesity activities in 3T3-L1 pre-adipocytes. Also, to improve anti-obesityeffects, research using 3T3-L1 pre-adipocytes cells is crucial. The anti-obesity effect of 70% ethanol extract from Torilis Japonica Decandolle on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated by suppressing adipocyte differentiation and lipid accumulation with Oil Red O assay, and western blot analysis. Compared to the control, 70% ethanol extract of Torilis Japonica Decandolle was significantly inhibited adipocyte differentiation and intracellular triglyceride (TG) level at a concentration of $100{\mu}g/ml$. To determine the mechanism of reduction in TG content, we determined the level of protein expression of obesity-related proteins, such as peroxisome-proliferatorsactivated-receptor-${\gamma}$ ($PPAR{\gamma}$) and CCAAT enhancer-binding-proteins-${\alpha}$ ($C/EBP{\alpha}$), and Acetyl-CoA carboxylase (ACC) phosphorylation. As a results, 70% ethanol extract of Torilis Japonica Decandolle significantly decreased protein expression of $PPAR{\gamma}$, $C/EBP{\alpha}$ and ACC phosphorylation. These results indicate that 70% ethanol extract of Torilis Japonica Decandolle is the most effective candidate for preventing obesity. However further studies will be needed to identify the active compounds that confer the anti-obesity activity of Torilis Japonica Decandolle.

The role of Fatty acid binding protein 5 (Fabp5) in fatty acid partitioning in the liver (간에서 지방산 분할에 대한 지방산결합 단백질 5의 역할)

  • Park, Jae-Seung
    • Journal of Digital Convergence
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    • v.17 no.8
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    • pp.283-291
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    • 2019
  • The aim of investigated the role of FABP5 in the hepatic lipogenesis and lipid metabolisms. Mice were overexpressed and silenced liver FABP5 using virus particles. Mice were fed a Western-type diet or regular chow for 1week and then sacrificed mouse after 24hr fasted. Liver homogenates were used for protein analysis by Western blot and mRNA levels by RT-PCR. Hepatic and serum lipids were analysed by thin-layer chromatography. Mice fed a Western-type or high saturated fat diet revealed large increases in FABP5 expression. However, FABP5 mRNA levels were drastically reduced under fasted. Hepatic TG was significantly increased FABP5-OEAV mice, but a significantly decreased hepatic free cholesterol under fed. The discovered a substantial decrease in hepatic TG mass with FABP5 silencing. In these data, presented evidence for an important role of FABP5 in hepatic lipogenesis and hepatic TG storage. FABP5 may also be a potential target in the treatment of NAFLD, metabolic syndrome, and obesity. Furthermore, studies to which transcription factors are involved in FABP5 expression and regulation.