• 제목/요약/키워드: Lipase assay

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Inhibitory Effects of 14 Plants from Mongolia and Myanmar on Lipid Accumulation in 3T3-L1 and HepG2 Cells (몽골과 미얀마 식물 14종의 3T3-L1 및 HepG2 세포에서 지질 축적 억제효과)

  • Kim, SukJin;Kim, Gun-Hee
    • Journal of the Korean Society of Food Culture
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    • v.36 no.1
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    • pp.130-142
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    • 2021
  • This study examined the antioxidative and lipid accumulation inhibitory effects of 14 plants from Mongolia and Myanmar on 3T3-L1 and HepG2 cells. The total phenolic and flavonoid contents (TPC and TFC) of 14 plant extracts were measured, and the antioxidative activities were analyzed using DPPH, ABTS, FRAP, and ORAC. After measuring the pancreatic lipase levels and performing the thiobarbituric acid assay, the degree of lipid accumulation was determined by lipid (Oil Red O) staining and triglyceride assay in 3T3-L1 and HepG2 cells. M. paniculate (259.43 mgGAE/g) and C. benghalensis (130.78 mgNAE/g) had the highest TPC and TFC, respectively, among the 14 plants. R. acicularis Lindl. had the highest antioxidant activity in DPPH. The ABTS, FRAP, and ORAC results showed that the antioxidant activity of 11 species was higher than that of the positive control. The pancreatic lipase inhibitory effect of C. angustifolium Scop. was reduced to 23.65% at 0.1 mg/mL, and the level of lipid peroxidation of C. abrorescens Lam. was 0.63 nmol/mg. Five selected plants inhibited the lipid accumulation and triglyceride content, respectively, in 3T3-L1 and HepG2 cells. These results provide scientific evidence for developing functional foods using 14 plants from Mongolia and Myanmar, which have antioxidant activities and lipid accumulation reduction effects.

Standardization of Pancreatin (판크레아틴의 규격 표준화 연구)

  • Shin, Ji-Eun;Yoon, Hae-Kyung;Kim, Dong-Hyun
    • Journal of Pharmaceutical Investigation
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    • v.33 no.4
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    • pp.273-279
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    • 2003
  • Pancreatin is a enzyme mixture breaking down carbohydrates, proteins and lipids. Most pancreatin used in Korea is imported from foreign countries. However, guideline of each country for pancreatin produced from each country is different. Therefore, guideline for pancreatin imported from several countries, such as Europe, Japan and America, it is standardized to control its quality. Assay of enzyme activity for pancreatin in KP is similar to tat in JP, but it is significantly different from those in FP ad in USP. We measured pancreatin digestive activities of 17 commercial products. Activity assay of digestive enzymes, starch- and lipid-digestive enzymes, for pancreatin by KP method (including JP) was difficult compared to those by FIP ad USP methods. Particularly, activity assays of starch- and lipid-digestive enzymes by KP method were mistakable, ad varied in diluted samples than those by FIP. However, activity assay of protein-digestive enzyme by KP method was similar to that by FIP. Starch-digestive enzyme activities of 17 commercial pancreatins by KP method were lower 0.079-fold compared to those by FIP method. Their protein-digestive enzyme activities by KP method were higher 75.7-fold than those by FIP method. Their lipid-digestive enzyme activities by KP method were lower 0.234-fold compared to those by FIP method.

Antioxidant and Antibacterial Activity of Caprylic Acid Vanillyl Ester Produced by Lipase-Mediated Transesterification

  • Kim, Jin Ju;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.317-326
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    • 2021
  • Vanillyl alcohol (VA), which is abundant in Vanilla bean, has strong antioxidant activity. However, the use of VA in the food and cosmetics industries is limited, due to its low solubility in emulsion or organic solvents. Meanwhile, medium chain fatty acids and medium chain monoglycerides have antibacterial activity. We synthesized butyric acid vanillyl ester (BAVE) or caprylic acid vanillyl ester (CAVE) from VA with tributyrin or tricaprylin through transesterification reaction using immobilized lipases. BAVE and CAVE scavenged 2,2-diphenyl-1-picrylhydrazyl radicals in organic solvents. In addition, BAVE and CAVE decreased the production rate of conjugated diene and triene in the menhaden oil-in-water emulsion system. While BAVE showed no antibacterial activity, CAVE showed antibacterial activity against food spoilage bacteria, including Bacillus coagulans. In this study, the antibacterial activity of vanillyl ester with medium chain fatty acid was first revealed. Zeta potential measurements confirmed that BAVE and CAVE were inserted into B. coagulans membrane. In addition, the propidium iodide uptake assay and fluorescent microscopy showed that CAVE increased B. coagulans membrane permeability. Therefore, CAVE is expected to play an important role in the food and cosmetics industries as a bi-functional material with both antioxidant and antibacterial activities.

Study of Lipoprotein Lipase Inhibitory Activity of Anti-obesity Herb Extracts (항비만소재의 lipoprotein lipase 억제 작용 연구)

  • Lee, Sung Mee;Kang, Yun Hwan;Kim, Kyoung Kon;Kim, Tae Woo;Choe, Myeon
    • Korean Journal of Food Science and Technology
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    • v.47 no.2
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    • pp.246-253
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    • 2015
  • In this study, we evaluated the lipoprotein lipase (LPL) inhibitory activity of 11 water extracts derived from Cinnamomum cassia Blume, Sarcodon aspratus, Cordyceps militaris, Crataegus pinnatifida Bunge, Corni fructus, Allium cepa, Coix lacryma-jobi, Plantago asiatica L., Lentinus edodes, Rosa rugosa, and Foeniculum fructus. The results of the LPL secretion and activity assay showed Sarcodon aspratus (NE) extract have an LPL secretion inhibitory acitivity. The cause of reduction in LPL secretion after NE treatment was investigated using molecular biology methods. NE treatment affected the LPL content in cells, but did not affect LPL mRNA expression. It also increased the mRNA expression level of sortilin-related receptor LDLR class A (SorLA), a receptor that induces endocytosis and intracellular trafficking of LPL. Finally, cell fractionation revealed that NE treatment induced the expression of CCAAT-enhancer-binding protein beta ($C/EBP{\beta}$), a SorLA transcription factor, in the nuclei of 3T3-L1 adipocytes. These results show that NE's anti-obesity effect involves inhibition of LPL secretion through $C/EBP{\beta}$-mediated induction of SorLA expression.

Evaluation of biological activities of plasma-treated phloridzin (플라즈마 처리 phloridzin 반응물의 생리활성 평가)

  • Jeong, Gyeong Han;Kim, Tae Hoon
    • Food Science and Preservation
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    • v.24 no.3
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    • pp.483-489
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    • 2017
  • Phloridzin is a predominant member of the chemical class of dihydrochalcones and mainly found in apple. The biological activity of phloridzin treated with dielectric barrier discharge (DBD) plasma was evaluated to investigate whether exposure to plasma can be used as a tools to enhance the biological activity of natural resources. DBD plasma treatment of phloridzin was carried out for three different exposure times. The antioxidant effects of degraded phloridzin for different reaction time were evaluated via radical scavenging assay using DPPH radical. In addition, the anti-diabetic and anti-obesity properties of the degraded phloridzin were measured based on ${\alpha}$-glucosidase and pancreatic lipase inhibitory activities. Phloridzin treated for 60 min showed significantly higher radical scavenging, ${\alpha}$-glucosidase and pancreatic lipase inhibitory activities compared to the tested positive controls. Degradation of phloridizin induced by DBD plasma might be responsible for enhancing the biological activity of phloridizin.

Protective Effects of Ethanol Extract from Aster Yomena on Acute Pancreatitis (쑥부쟁이 에탄올 추출물의 급성췌장염 보호 효과)

  • Seo, Sang Wan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.33 no.2
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    • pp.109-115
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    • 2019
  • Aster Yomena (AY) has been used in traditional medicine to treat diseases such as obesity, hyperlipidemia, atherosclerosis, diabetes and osteoarthritis. However, protective effect of AY on acute pancreatitis (AP) has not been reported. The present study examined the anti-inflammatory effects of an ethanol extract of AY on cerulein-induced AP. AP was induced in mice by intraperitoneally injecting cerulein ($50{\mu}g/kg$) hourly for 6 times. 70% ethanol extract of AY (0.1, 0.2, and 0.5 g/kg) was orally administered for 1 week before acute pancreatitis induction. The mouse was killed at 6 hours after the final cerulein injection. The pancreas and lung were rapidly removed for histological examination and myeloperoxidase (MPO) assay. Blood samples were taken to determine serum amylase and lipase activity. In addition real-time reverse transcription-polymerase chain reaction (RT-PCR) was also performed to investigate mRNA expression of proinflammatory cytokines such as $TNF-{\alpha}$. $IL-1{\beta}$, and IL-6. Administration of AY significantly ameliorated pancreatic weight to body weight ratio, histological damages and MPO activity during AP. In addition, AY inhibited the serum amylase and lipase activity during AP. Also, mRNA expression of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 were inhibited by AY against AP. Our results revealed that pre-treatment of AY reduces the severity of cerulein-induced AP. Therefore, AY may have a protective effect drug against AP.

Lipase-Inhibitory and Anti-Oxidative Activity of the Methanol Extract and the Powder of Phellinus linteus (상황버섯 자실체 메탄올 추출물과 분말의 지방소화효소 억제 및 항산화 활성)

  • Kim, Ji-Hyun;Son, In-Suk;Kim, Jong-Sang;Kim, Ki-Hoon;Kwon, Chong-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.2
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    • pp.154-161
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    • 2008
  • Phellinus linteus (PL) has been known to exhibit potent biological activity. The present study was designed to investigate lipase-inhibitory and anti-oxidative activity of the methanol extract and the powder of PL fruiting body. The methanol extract of PL appeared to have the inhibitory activity against pancreatic lipase with an $IC_{50}$ value of $36.3\;{\mu}g/mL$, and the scavenging activity of DPPH radical with an $IC_{50}$ value of $20.1\;{\mu}g/mL$, which was similar to that of vitamin C ($IC_{50}\;18.3\;{\mu}g/mL$). To investigate the lipase-inhibitory and anti-oxidative effect of PL on animal, Sprague-Dawley rats were fed with high-fat diet supplemented with either 2% or 5% PL powder for 8 weeks. Total food intake was significantly increased, but body weight was not changed by PL powder supplementation. However, fecal fat excretion of the experimental groups fed with the PL powder were higher than that of the control group. PL powder showed a decrease in the plasma total cholesterol, LDL-cholesterol, and the hepatic total cholesterol levels. The anti-oxidative enzyme activities were also affected by PL supplementation. Glutathione peroxidase (GSH-Px) in the plasma and liver were significantly increased by 98% and 46% in the 2% PL group, and 99% and 32% in the 5% PL group, respectively. Total superoxide dismutase (T-SOD) activity was not affected by PL supplementation. DNA damage was measured by the comet assay in the lymphocytes collected after 2 weeks, 4 weeks, and 8 weeks of feeding PL supplemented diet. Lymphocyte DNA damage was decreased in the PL supplemented group. Furthermore, PL feeding enhanced the resistance to lymphocyte DNA damage caused by an oxidant challenge with $H_2O_2$.

Improvement of anti-oxidant and anti-inflammatory activities of aleurone layer extracts of black rice (Oryza sativa L.) by enzyme treatment (효소 처리에 의한 흑미 호분 추출물의 산화방지와 항염증 활성 증진)

  • Lee, Mi Kyoung;Ryu, Soo In;Lee, Min Ho
    • Korean Journal of Food Science and Technology
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    • v.50 no.5
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    • pp.528-534
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    • 2018
  • The current study investigated the effects of enzyme treatment on black rice (Oryza sativa L.) aleurone layer extracts. Different enzymes (lipase, lecitase ultra and lipopan 50BG) were used to test anti-oxidant and anti-inflammatory activities in vitro. The antioxidant activities of enzyme treated or non-enzyme treated extracts of black rice bran were evaluated via 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity. Lipase treated extracts of black rice bran showed higher antioxidant activity compared to that of non-enzyme treated extracts. Anti-inflammatory activities of enzyme treated black rice bran extracts on nitrite production and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) secretion, were tested using a nitric oxide (NO) colorimetric assay kit and an enzyme-linked immunosorbent assay (ELISA) kit. The ethanolic extract of enzyme treated black rice bran decreased the levels of nitric oxide production and pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$, in a lipopolysaccharidestimulated RAW 264.7 cell culture. These findings indicate that enhanced anti-oxidant and anti-inflammation activities of the ethanolic extracts of enzyme treated black rice (Oryza sativa L.) aleurone layers, may be attributed to molecular conversion of ingredients in enzyme catalyzed reactions.

Anti-adipogenic Effect of Undaria pinnatifida Extracts by Ethanol in 3T3-L1 Adipocytes (미역 에탄올 추출물이 지방세포 형성과정에 미치는 영향)

  • Kim, Hye-Jin;Kang, Chang-Han;Kim, Sung-Koo
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1052-1056
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    • 2012
  • Undaria pinnatifada has been used as a natural diet food with few calories and as a source of iodine. Even though U. pinnatifida has been regarded as a diet food, the mechanisms of its inhibitory effects on adipocyte differentiation and the accumulation of fat in adipocytes are poorly understood. In this study, the effect and mechanism of U. pinnatifida ethanol extract on 3T3-L1 differentiation into adipocytes were investigated. The effects of U. pinnatifida ethanol extract on cell viability and the anti-adipogenic effect were investigated via MTT assay, Oil red O staining, RT-PCR, and western blot. The U. pinnatifida ethanol extract did not show toxicity up to a concentration of 50 ${\mu}g/ml$. The addition of U. pinnatifida ethanol extract decreased triglyceride contents by 40% when 50 ${\mu}g/ml$ of U. pinnatifida ethanol extract was added during 3T3-L1 differentiation and adipocyte triglyceride formation. The transcription and expression of peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), leptin, and hormone-sensitive lipase (HSL) as adipocyte-specific proteins were determined by RT-PCR and western blot. The overexpression of $PPAR{\gamma}$ could accelerate adipocyte differentiation. Also, leptin was secreted for triglyceride accumulation in the adipocytes and the increase of adipocyte cell size. Thus, $PPAR{\gamma}$ and leptin were used as indicators of obesity. $PPAR{\gamma}$ and leptin were repressed by the increased addition of U. pinnatifida ethanol extract. This indicates that U. pinnatifida was effective as an anti-obesity agent by repressing the differentiation of 3T3-L1 into adipocytes and inhibiting triglyceride formation in adipocytes.

Evaluation of free radical scavenging and pancreatic lipase inhibitory effects of Aquilaria agallocha extracts (침향 추출물의 라디칼 및 췌장 지방분해 효소저해 활성 평가)

  • Lee, Ha Yeong;Lee, In-Chul;Kwak, Jae Hoon;Kim, Tae Hoon
    • Food Science and Preservation
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    • v.22 no.3
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    • pp.437-442
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    • 2015
  • In a continuing screening of selected medicinal plants native to South Korea, the antioxidant and pancreatic lipase inhibitory activities of an aqueous methanolic extract from the heartwood of Aquilaria agallocha were investigated. Eighty percent of the methanolic extract of A. agallocha was further divided into $CH_2Cl_2$, EtOAc and n-BuOH in order to yield four solvent-soluble portions, namely $CH_2Cl_2$-soluble, EtOAc-soluble, n-BuOH-soluble and $H_2O$ residue. The antioxidant properties were evaluated by employing radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) ($ABTS^+$) radicals, while the anti-obesity efficacy of A. agallocha extracts and solvent-soluble portions were tested by porcine pancreatic lipase assay. All tested samples showed dose-dependent radical scavenging and pancreatic lipase inhibitory activities. Among the tested extracts and solvent-soluble portions, the $CH_2Cl_2$-soluble portion showed much higher radical scavenging activity and pancreatic lipase inhibitory properties when compared with other solvent-soluble portions. This result suggested that there was a significant relationship between the total phenolic content and biological efficacies, and A. agallocha extract might be considered as a new potential source of natural antioxidants and as a pancreatic lipase inhibitory source. A more systematic investigation of this biomass will be performed for further investigation of activity against antioxidative and anti-obesity effects.