• Title/Summary/Keyword: Lipase assay

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Association of Backfat Thickness with Postheparin Lipoprotein Lipase Activity and Very Low Density Lipoprotein-Subfractions in Growing Pigs

  • Loh, T.C.;Lean, I.J.;Dodds, P.F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.11
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    • pp.1592-1597
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    • 2001
  • Sixteen pigs from 2 distinct genetic lines (LGAH and VFIL) obtained after eight generations of divergent selection for high (H) and low (L) lean tissue growth rate with ad-libitum feeding (LGA) and voluntary feed intake (VF1), respectively, were used in this study. The objectives of this investigation were to establish appropriate working conditions for the postheparin plasma lipoprotein lipase (LPL) assay and to study relationships between fat deposition and plasma lipids, very low density lipoprotein (VLDL) lipids, VLDL-subfractions and postheparin plasma LPL activity in growing pigs. Four preliminary experiments were performed to determine the appropriate working conditions for the postheparin plasma LPL assays. Postheparin plasma preincubated with SDS (20-50 mM) at $26^{\circ}C$ for 45 minutes inhibited hepatic lipase activity. A total of $2{\mu}l$ VLDL/assay produced maximum stimulation of LPL activity. Postheparin plasma protein and increasing incubation time contributed an optimum response. LGAH pigs had a significantly higher proportion subtraction 2 than VFIL pigs. No differences were observed in postheparin plasma LPL activity and backfat thickness for two lines of pigs. There were positive correlations between backfat thickness and proportion of subtractions 2 and postheparin plasma LPL activity but the results were not statistically significant. Backfat thickness was not statistically correlated with proportion of subtraction 2 and postheparin plasma LPL activity in a multiple regression analysis. It is believed that the apolipoprotein E, which is present in higher quantities in VLDL-subfraction 2 plays an important role for clearing VLDL triacylglycerol into adipose tissue. LPL activity of pigs can be measured by using postheparin plasma technique. If the relationships of backfat thickness and VLDL-subfraction 2 and postheparin plasma LPL activity can be established, it suggests that these parameters could be used as indicators in selection programmes. Further experiments need to be conducted by using larger sample size and different breed of pigs with greater differences in backfat thicknesses to confirm these trends.

Anti-Oxidative and Anti-Obesity Effects of Amomum Cardamomum L. Extract (백두구 추출물의 항산화 및 항비만 효과)

  • Park, Jung Ae;Jin, Kyong-Suk;Lee, Ji Young;Kwon, Hyun Ju;Kim, Byung Woo
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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    • pp.249-257
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    • 2014
  • In this study, the anti-oxidative and anti-obesity activities of Amomum cardamomum L. methanol extract (ACME) were evaluated using DPPH radical scavenging activity assay, pancreatic lipase enzyme inhibition assay, and the cell culture model system. ACME exhibited DPPH radical scavenging activities dose-dependently, with $IC_{50}$ of DPPH radical scavenging activities of ACME being $25.15{\mu}g/ml$. Furthermore, ACME effectively suppressed pancreatic lipase enzyme activity dose-dependently. ACME also significantly suppressed adipocyte differentiation, lipid accumulation, triglyceride (TG) contents, and triggered lipolysis activity on 3T3-L1 preadipocytes in a dose-dependent manner, without cytotoxicity. Their anti-obesity effect was modulated by the cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins ${\alpha}$ ($C/EBP{\alpha}$), $C/EBP{\beta}$ and the peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) gene and protein expressions. Taken together, these results provide an important new insight that A. cardamomum L. possesses anti-oxidative and anti-obesity activities such as pancreatic lipase inhibition, anti-adipogenic, and lipolysis effects. There is therefore potential for its use as a promising component in the field of nutraceuticals and the identification of the active compounds that confer the anti-oxidative and anti-obesity activities of ACME might be an appropriate next step.

Inhibitory Effects of Marine Algae Extract on Adipocyte Differentiation and Pancreatic Lipase Activity

  • Kim, Eun-Sil;Lee, Kyoung-Jin;Oh, Kyoung-Hee;Ahn, Jong-Hoon;Kim, Seon-Beom;Liu, Qing;Hwang, Bang-Yeon;Lee, Mi-Kyeong
    • Natural Product Sciences
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    • v.18 no.3
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    • pp.153-157
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    • 2012
  • Obesity, which is characterized by excessive fat accumulation in adipose tissues, occurs by fat absorption by lipase and sequential fat accumulation in adipocyte through adipocyte differentiation. Thus, inhibition of pancreatic lipase activity and adipocyte differentiation would be crucial for the prevention and progression of obesity. In the present study, we attempted to evaluate anti-adipogenic activity of several algae extracts employing preadipocytes cell line, 3T3-L1 as an in vitro assay system. The effects on pancreatic lipase activity in vitro were also evaluated. Total methanolic extracts of Cladophora wrightiana and Costaria costata showed significant inhibitory activity on adipocyte differentiation as assessed by measuring fat accumulation using Oil Red O staining. Related to pancreatic lipase, C. wrightiana and Padina arborescens showed significant inhibition. Further fractionation of C. wrightiana, which showed the most potent activity, suggested that $CHCl_3$ and n-BuOH fraction are responsible for adipocyte differentiation inhibition, whereas n-BuOH and $H_2O$ fraction for pancreatic lipase inhibition. Our study also demonstrated that n-BuOH fraction was effective both in early and middle stage of differentiation whereas $CHCl_3$ fraction was effective only in early stage of differentiation. Taken together, algae might be new candidates in the development of obesity treatment.

Characterization of $\alpha$-amylase, Total Alkaline Pretense, Trypsin and Triacylglycerol-lipase Activity of the Euryhaline Rotifer Brachionus rotundiformis (해수산 rotifer, Brachionus rotundiformis의 $\alpha$-amylase, total alkaline Protease, trypsin 및 triacylglycerol-lipase 활성 특성)

  • Kwon O-Nam;Park Heum-Gi
    • Journal of Aquaculture
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    • v.18 no.4
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    • pp.245-251
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    • 2005
  • This study was investigated the condition of their maximum activity to assay the enzymes of rotifer, Brachionus rotundiformis의 $\alpha$-amylase, total alkaline Protease, trypsin and TG-lipase activities of rotifer were higher and more sensitive in phosphate-NaOH buffer than Tris-HCl buffer. $\alpha$-amylase, trypsin and TG-lipase activities were appeared the maximum at pH 8.0, and total alkaline protease activity showed the maximum activity at pH 7.0. $\alpha$-amylase activity showed the highest activity at $40^{\circ}C$, and total alkaline protease and trypsin activities were assayed the highest at $55{\~}60^{\circ}C$. However, TG-lipase activity was appeared the highest at $25{\~}30^{\circ}C$. The optimum substrate concentration of enzyme activity of a-amylase, total alkaline protease, rypsin and TG-lipase were $3.5\%$ starch, $\0.6%$ azo-casein, $87.5{\mu}M$ BApNA and 81.2 mM olive oil, respectively. The optimum reaction time of enzyme activity of $\alpha$-amylase, total alkaline protease, trypsin and TG-lipase were increased up to 40, 60, 30 and 25 min., respectively. The data obtained in this study could be used for the digestive enzyme research of rotifer, B. rotundiformis.

Antioxidant and pancreatic lipase inhibitory activities of Anemarrhena asphodeloides (지모 추출물의 항산화 및 pancreatic lipase 저해 활성 평가)

  • Kwon, O Jun;Lee, Ha Yeong;Kim, Tae Hoon;Kim, Se Gie
    • Food Science and Preservation
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    • v.21 no.3
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    • pp.421-426
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    • 2014
  • In this study, the antioxidant and pancreatic lipase inhibitory activities of aqueous methanolic (70% methanol) extract from the roots of Anemarrhena asphodeloides were investigated. The extracts of four solvent fractions (the n-hexane layer, EtOAc layer, n-BuOH layer, and $H_2O$ layer) of the 70% methanol extract were also investigated. Furthermore, the total phenolic content was quantified using a spectrophotometric method. All the tested samples showed dose-dependent radical scavenging and pancreatic lipase inhibitory activities. In particular, the pancreatic lipase inhibitory activity of the ethyl acetate soluble portion (the EtOAc layer) from the rhizomes of the A. asphodeloides was higher than that of the other solvent-soluble portions. The antioxidant property of the extracts was evaluated using radical scavenging assays with DPPH and $ABTS^+$ radicals. 1000 mg/ml of the n-BuOH layer extract showed 91.2% DPPH radical scavenging activity. The EtOAc layer extract and the n-BuOH layer extract showed $IC_{50}=20.5{\pm}1.7mg/ml$ and $IC_{50}=50.5{\pm}0.7mg/ml$ $ABTS^+$ radical scavenging activities, respectively. The anti-obesity efficacy of the A. asphodeloides extract was tested via porcine pancreatic lipase assay. A pancreatic lipase inhibitory activity ($IC_{50}$) of $31.3{\pm}0.1mg/ml$ was obtained from the EtOAc layer extract. These results suggest that A. asphodeloides can be considered a new potential source of natural antioxidant and anti-obesity agents.

Secondary Metabolites from Enzymatic Oxidation of Caffeic Acid with Pancreatic Lipase Inhibitory Activity (카페인산의 효소적 산화반응으로부터 췌장 지방분해효소 저해 물질의 분리)

  • Kim, Tae Hoon;Kim, Myoung Kwon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.12
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    • pp.1912-1917
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    • 2015
  • Pancreatic lipase is a potential therapeutic target for the treatment of diet-induced obesity in humans. As part of our continuing search for novel bioactive compounds, the convenient enzymatic transformation of caffeic acid into neolignans as well as related oxidized-products enhanced pancreatic lipase inhibitory activity. Enzymatic transformation of caffeic acid (1) using polyphenol oxidase originating from Korean pear yielded four oxidized metabolites, which were identified by different spectroscopic techniques ($^1H$,$^{13}C$ NMR, DEP/T, $^1H-^1H$ COSY, HMBC, HMQC, and NOESY). The anti-obesity efficacy of caffeic acid reactant was tested by in vitro porcine pancreatic lipase assay. All tested samples showed dose-dependent pancreatic lipase inhibitory activities. Four oxidative products including phellinsin A (2), caffeicinic acid (3), isocaffeicinic acid (4), and 7,8-erythro-caffeicin (5) were isolated and identified. The major metabolites (2~5) were evaluated for their pancreatic lipase inhibitory activity, and oxidized-products (2~3) improved potency against pancreatic lipase when compared to original caffeic acid. This result suggested that the neolignans isolated from oxidative transformation of caffeic acid might be beneficial in the treatment of obesity and relevant diseases, and the convenient enzymatic transformation by polyphenol oxidase may be a valuable method for structural modification and enhancement of activity.

Pancreatic Lipase Inhibitory and Antioxidant Activities of Zingiber officinale Extracts (생강 추출물의 pancreatic lipase 저해 및 항산화 활성)

  • Bae, Jong-Sup;Kim, Tae-Hoon
    • Food Science and Preservation
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    • v.18 no.3
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    • pp.390-396
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    • 2011
  • Ginger (Zingiber officinale) is a well-known herb that is widely consumed as spice for the flavoring of foods. As part of our continuing search for bioactive materials, the in vitro pancreatic lipase inhibition and antioxidant properties of an aqueous ethanolic extract of Z. officinale were investigated. The total phenolic content was determined using a spectrophotometric method. The antioxidant efficacies of the extract was studied with radical scavenging assays using DPPH and $ABTS^+$ radicals. Further more, the antiobesity effect of the extract was evaluated by porcine pancreatic lipase assay. In particularly, the pancreatic lipase inhibitory activity of the ethyl acetate (EtOAc)-soluble portion from Z. officinale was significantly higher than that of the other solvent-soluble portions. The results suggest that Z. officinale may have therapeutic potential that may be useful in development of an anti-obesity agent or its precursors.

Pancreatic Lipase Inhibitory and Antioxidant Activities of Polygonum aviculare Extract (마디풀 추출물의 Pancreatic Lipase 저해 및 항산화 활성 평가)

  • Kim, Tae-Hoon
    • Food Science and Preservation
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    • v.18 no.2
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    • pp.250-255
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    • 2011
  • As part of our continuing search for novel bioactive natural products, antioxidant and pancreatic lipase inhibitory activities of the aqueous ethanolic extract of Polygonum aviculare were investigated. In addition, the total phenolic content was quantified by a spectrophotometric method. The antioxidant properties of the extract were evaluated by radical scavenging assays such as DPPH and $ABTS^+$ radicals. The anti-obesity efficacy of P. aviculare extract was tested by porcine pancreatic lipase assay. All tested samples showed a dose-dependent pancreatic lipase inhibitory and radical scavenging activities. In terms of the $IC_{50}$ value, the ethyl acetate (EtOAc) soluble portion was most potent than other solvent soluble portions. The results suggested that P. aviculare could be considered as a new potential source of natural antioxidant and anti-obesity agent for food, nutraceutical, cosmetics industries.

Pancreatic Lipase Inhibitors Isolated from the Leaves of Cultivated Mountain Ginseng (Panax ginseng) (산양삼 잎으로부터 Pancreatic lipase 저해 활성물질의 분리)

  • Hong, Ju-Yeon;Shin, Seung-Ryeul;Bae, Man-Jong;Bae, Jong-Sup;Lee, In-Chul;Kwon, O-Jun;Jung, Ji-Wook;Kim, Yong-Han;Kim, Tae-Hoon
    • Food Science and Preservation
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    • v.17 no.5
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    • pp.727-732
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    • 2010
  • Activity-guided fractionation of an ethyl acetate (EtOAc)-soluble portion of an ethanolic extract from the leaves of cultivated mountain ginseng, using pancreatic lipase inhibition assay, led to the isolation and identification of three flavonoids of a previously described structure, kaempferol-3-O-sophoroside (I), kaempferol-3-O-${\beta}$-Dglucopyranoside (astragalin, II) and kaempferol (III). All compounds (I.III) showed pancreatic lipase inhibitory activities, with $IC_{50}$ values ranging from $20.3{\pm}2.2$ to $9.1{\pm}1.5$ ${\mu}M$, kaempferol (III) showed the most potent inhibitory activity with an $IC_{50}$ of $9.1{\pm}1.5$ ${\mu}M$. The level of activity may depend on the number of C-3 glucosyl group(s) linked to the kaempferol backbone, and the isolated compounds may have promise as pancreatic lipase inhibitors.

Detection of urinary trypsinogen-2 for diagnosis of canine acute pancreatitis (뇨중 trypsinogen-2 검출을 이용한 개의 급성 췌장염 진단)

  • Han, Hong-ryul;Hwang, Cheol-yong;Pak, Son-il;Oh, Tae-ho
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.951-960
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    • 1999
  • We performed this study to evaluate the potential clinical marker of urinary trypsinogen-2 together with amylase, lipase and urinary amylase creatinine clearance ratio (ACCR) for the diagnosis of acute pancreatitis in dogs. In the experiment on daily changing patterns of amylase, lipase and ACCR measurements in experimentally induced pancreatitis dogs, compared to values measured in pre-induction state, significant difference was seen in amylase until 5th day of induction, and for lipase significant difference was found during the 7th day of observation period (p < 0.05). No significant difference was found in ACCR for the study period (p > 0.05). On SDS-PAGE analysis of urine from experimentally induced pancreatitis dog, The 26kd band was markedly increased compared with that of normal state and that band was confirmed trypsinogen-2 using substrate interaction and isoelectric focusing assay after being eluted. When assessing the appearance of 26kd band on urine SDS-PAGE 87.1% (range: 50~100%) of experimentally induced pancreatitis dogs showed positive results, whereas no corresponding band was seen in dog without pancreatic disorders. With this result, determination of urinary trypsinogen-2 assay was found to have a high diagnostic value with a 70% of sensitivity and 100% of specificity as a routine test for pancreatitis, although the detection of trypsinogen-2 in urine can be varied on the progression stage of pancreatitis at the initial visit to animal clinic. We therefore suggest that the promising results in this study be used for the development of dipstick test for detecting acute pancreatitis in the future research.

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