• 제목/요약/키워드: Activity assay

검색결과 5,696건 처리시간 0.036초

Determination of the Kinetic Properties of Platy cod in D for the Inhibition of Pancreatic Lipase Using a 1, 2-Diglyceride- Based Colorimetric Assay

  • Zhao, Hai-Un;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • 제27권9호
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    • pp.968-972
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    • 2004
  • A 1, 2-diglyceride-based multi-step colorimetric assay to measure the pancreatic lipase activ-ity was applied for the determination of the kinetic profiles of the lipase inhibition with a slight modification and the validity verification. With this assay method, our study revealed that platy-codin D, one of major constituents of Platycodi Radix, inhibits the pancreatic lipase activity in a competitive type, with the value of $K_1$ being 0.18${\pm}$0.02 mM. In addition, PO has affected the val-ues of $K_{m}$, app/ and $K_{cat}$/$K_{m}$ in a dose-dependent manner. The results shed a meaningful light on how PO mediates lipid metabolism in the intestinal tracts. On the other hand, since the revised assay is sensitive, rapid, and does not affect the accuracy to the kinetic properties, it is applica-ble not only to evaluation of the kinetic properties of the pancreatic lipase, but also to high-throughput screening of pancreatic lipase activity.

In vitro Angiogenic Activity of Aloe vera Gel on Calf Pulmonary Artery Endothelial (CPAE) Cells

  • Lee, Myoung-Jin;Lee, Ok-Hee;Yoon, Soo-Hong;Lee, Seung-Ki;Chung, Myung-Hee;Park, Young-In;Sung, Chung-Ki;Choi, Jae-Sue;Kim, Kyu-Won
    • Archives of Pharmacal Research
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    • 제21권3호
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    • pp.260-265
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    • 1998
  • Angiogenic activity of Aloe vera gel was investigated by in vitro assay. We obtained the most active fraction from dichloromethane extract of Aloe vera gel by partitioning between hexane and 90% aqueous methanol. The most active fraction (F3) increased the proliferation of calf pulmonary artery endothelial (CPAE) cells. In addition, F3 fraction induced CPAE cells to invade type I collagen gel and form capillary-like tube through in vitro angiogenesis assay, and increased the invasion of CPAE cells into matrigel through in vitro invasion assay. Furthermore, the effect on the MRNA expression of proteolytic enzymes which are key participants in the regulation of extracellular matrix degradation was investigated by northern blot analysis. F3 fraction enhanced mRNA expression of urokinase-type plasminogen activator (u-PA), matrix metalloproteinase-2 (MMP-2), and membrane-type MMP (MT-MMP) in CPAE cells whereas the expression of plasminogen activator inhibitory (PAl-1) mRNA was not changed.

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천연 Quercitrin의 항허피스바이러스작용과 Nucleoside계 항허피스바이러스제와의 병용효과 (Antiherpetic Activities of Natural Quercitrin Alone and in Combinations with Nucleoside Antiherpetic Agents)

  • 김영소;어성국;김홍진;이도익;김기호;한성순
    • Biomolecules & Therapeutics
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    • 제7권2호
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    • pp.158-163
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    • 1999
  • In order to find less toxic antiherpetic agents, antiviral activities of quercitrin against two strains of pathogenic viruses such as herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) were determined in Vero cells using plaque reduction assay in vitro. Quercitrin showed a concentration-dependent decrease in plaque formation of HSV-1 and HSV-2. It also exhibited more potent antiherpetic activity on HSV-1 with 50% effective concentration (EC$_{50}$) of 20.4 $\mu$g/ml than on HSV-2 with EC$_{50}$ of 30.4 $\mu$g/ml. The combined antiherpetic effects of quercitrin with nucleoside antiherpetic agents, acyclovir and vidarabine, were examined on the multiplication of these two strains of herpesviruses in Vero cells by the combination assay. The results of combination assay were evaluated by the combination index (CI) that was calculated by the multiple drug effect analysis. The combinations of quercitrin with acyclovir and vidarabine on HSV-1 showed more potent synergism with CI values of 0.27-0.81 for 50%, 70%, 90% effective levels than those on HSV-2 with CI values of 1.03~2.20..20.

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사물탕, 사군자탕, 팔물탕, 십전대보탕의 약리 활성 비교 연구 (Immuno stimulatory activities of Samul-tang, Sagunja-tang, Pamul-tang and Sipjeondaebo-tang in vitro)

  • 이호영;하혜경;정다영;이남헌;신현규
    • 동의신경정신과학회지
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    • 제21권4호
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    • pp.41-51
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    • 2010
  • Objectives : Samul-tang(SM). Sagunja-tang(SG). Pamul-tang(PM) and Sipjeondaebo-tang(SJ) was used many diseases such as sterility. menstrual disorder. general prostration. recruitment in Korea. We investigated the immune stimulatory activities of SG. SM. PM and SJ in in vitro. Methods: For comparision for effective of SM. SG. PM and SJ. this study examined anti-inflammation(NO. PGE2 assay). anti-oxidation(DPPH assay) and immune response in in vitro assay. For immune response activities. this study used NO synthesis on RAW 264.7 cells, splenocyte proliferation and cytokine assay(IL-2, IL-4) in splenocyte. Results: The results showed that SG. SM. PM and SJ were no significant effect anti-oxidation and anti-inflammatory effects. For immune response. they showed the splenocyte proliferation and macrophage proliferation. We confirmed that they synthesised NO a dose-dependent manner significantly and secreted the IL-4. Conclusions : These results suggested the SG, SM, PM and SJ had immune stimulatory activity. A efficacy of immune response in them had produced similar results.

In vitro antioxidant and free radical scavenging activities of stem extract of Euphorbia trigona Miller

  • Salar, Raj Kumar;Sharma, Pooja;Purewal, Sukhvinder Singh
    • 셀메드
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    • 제5권2호
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    • pp.14.1-14.6
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    • 2015
  • Antioxidative and free radical scavenging properties of different stem extracts of Euphorbia trigona were evaluated and correlated with its total phenolic content. Aqueous, acetone and methanolic extracts of shade dried stem were obtained and were concentrated in vacuo. The antioxidant and free radical scavenging activities of stem extracts was determined by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging assay, reducing power assay, deoxyribose degradation assay and $Fe^{2+}$ chelating assay. Total phenolic contents (TPC) were evaluated using Folin-Ciocalteu reagent. The results confirmed that the plant is a rich source of polyphenolic compounds which are invariably higher compared to other herbs. All extracts showed TPC in the range of 146.6 - 168.6 mg/g gallic acid equivalents at $300{\mu}g/ml$ of extract. Among the three extracts ME showed highest scavenging activity as evidenced by maximum scavenging of DPPH (83.2%), $OH{\bullet}$ radicals (94.81%), $Fe^{2+}$ chelating activity (88.59%) and a high reducing power 0.623 at $300{\mu}g/ml$. Our results demonstrate that Euphorbia trigona, an unexplored xerophytic plant could be potential source of natural antioxidants and phytotherapeutic agents. The plant possess invariably high amount of polyphenolic compounds with a broad spectrum of antioxidant properties and could be further used for food, feed and pharmaceutical applications.

천연 나린게닌의 항허피스바이러스작용과 아시클로버 및 비다라빈과의 병용효과 (Antiherpetic Activities of Natural Naringenin Alone and in Combinations with Acyclovir and Vidarabine)

  • 이지현;김영소;이종길;이혁구;한성순
    • 약학회지
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    • 제43권1호
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    • pp.77-84
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    • 1999
  • To search for less toxic antiherpetic agents, the inhibitory effects of natural naringenin on the plaque formation of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in Vero cells were examined by the plaque reduction assay in vitro. Naringenin inhibited plaque formations of HSV-l and HSV-2 in a dose dependent manner. It also exhibited more potent antiherpetic activity on HSV-l with selectivity index (SI) of 19.1 than on HSV-2 with SI of 5.7 The combined antiherpetic effects of naringenin with nucleoside antiherpetic agents, acyclovir and vidarabine, were examined on the multiplication of these two strains of herpesviruses in Vero cells by the combination assay. The results of combination assay were evaluated by the combination index (CI) that was calculated by the multiple drug effect analysis. The combinations of naringenin with acyclovir on HSV-l and HSV-2 showed more potent synergism with CI values of 0.28∼0.81 for 50%, 70%, 90% effective levels than those with vidarabine with CI values of 0.86-3.28.

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한약조성물 KCNS-001이 자유라디칼과 염증매개인자에 미치는 영향 (Anti-inflammatory Effects of Herbal Formula KCNS-001 for Mitigating Atopic Dermatitis)

  • 이정복;최재환;방옥선;유영법
    • 대한본초학회지
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    • 제24권3호
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    • pp.97-102
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    • 2009
  • Objectives : We determined the anti-inflammatory activity of KCNS-001 that is a herbal formula including 6 medicinal plants and that are used to mitigate atopic dermatitis in oriental medicine. Methods : To evaluate anti-inflammatory effect of KCNS-001, we measured the production of reactive oxygen species (ROS), nitric oxide (NO) and cyclooxygenase-2 (COX-2) in LPS-activated Raw 264.7 cells. Cell viability was determined by MTT assay. The concentrations of ROS and relative level of NO were measured with DPPH assay and Griess reagent, respectively. COX-2 and TNF-$\alpha$ were detected by enzyme immuno assay (EIA) and enzyme-linked immunosorbent assay (ELISA). Results : ROS and NO production were reduced by KCNS-001 in a dose-dependent manner. KCNS-001 significantly inhibited activity of COX-2 and suppressed the release of tumor necrosis factor-alpha (TNF-$\alpha$). Conclusions : These results indicate that the KCNS-001 may have an anti-inflammatory agent for the treatment of various inflammatory disease.

TOSC 법을 이용한 당알코올의 항산화 활성 평가 (Evaluation of Antioxidant Activity of Sugar Alcohols Using TOSC (Total Oxy-radical Scavenging Capacity) Assay)

  • 강건욱;곽상훈;윤세영;김상겸
    • Toxicological Research
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    • 제23권2호
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    • pp.143-150
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    • 2007
  • Although animal and epidemiological studies have suggested oxidative stress as an etiological factor in pathogenesis including cancer, inflammation, sepsis, fibrosis, cardiovascularlneurodegenerative diseases and aging-related disorders, conflicting results have been obtained in clinical trial with antioxidants. The reason for this discrepancy remains unknown but may be due, in part, to the lack of a validated assay system for evaluating antioxidant capacity. The antioxidant activity of a series of sugar alcohols against peroxyl radicals, hydroxyl radicals and peroxynitrites was determined by the total oxy-radical scavenging capacity (TOSC) assay and cell-based assay using H4IIE cells. Specific TOSC values calculated from the slope of the linear regression for erythritol, xylitol, sorbitol or mannitol against peroxyl radicals was $2.1{\pm}0.2,\;3.7{\pm}0.3,\;9.1{\pm}0.3$ or $8.7{\pm}1.1$ TOSC/mM, respectively. Specific TOSC values for erythritol, xylitol, sorbitol or mannitol against peroxynitrite was $1.9{\pm}0.3,\;3.9{\pm}0.4,\;7.8{\pm}0.7$ or $7.7{\pm}0.5$ TOSC/mM, respectively. These results suggest that oxy-radical scavenging capacity is dependent on the number of aliphatic hydroxyl group in sugar alcohols of monosaccharide. Tert-butylhydroperoxide (t-BHP)-induced cell toxicity determined by MTT assay was marginally attenuated by 10 mM erythritol, but completely inhibited by 10 mM xylitol, 2 mM sorbitol or 0.75 mM maltitol, a disaccharide alcohol. Oxidative stress markers, such as glutathione (GSH) and malondial-dehyde (MDA) levels, were measured in t-BHP-treated cells using HPLC equipped with a fluorescence detector and a reverse phase column. Erythritol did not change the levels of GSH and MDA in H411E cells treated with t-BHP. The t-BHP-induced changes in cellular GSH and MDA levels were ameliorated by 10 mM xylitol and completely blocked by 10 mM sorbitol and maltitol. These results indicate that sugar alcohols protect cells against oxidative stress via scavenging oxy-radical and suggest that TOSC assay in conjunction with cell-based assay is a valid method for evaluating antioxidant capacity of natural and synthetic chemicals.

Study on Physiologically Active Compounds and Antioxidant Activity of Korean Yam (Dioscorea batatas DECNE.)

  • Duan, Yishan;Kim, Gyeong-Hwuii;Kim, Han-Soo
    • 한국응용과학기술학회지
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    • 제33권2호
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    • pp.342-351
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    • 2016
  • The bioactive compound and antioxidant property of Korean yam (Dioscorea batatas DECNE.) were studied using in vitro methods. Yam available in Korea was analyzed for lycopene, chlorophyll a, b, tannin, phytic acid and total saponin contents. 70% Methanol, 70% ethanol and chloroform-methanol mixture (CM, 2:1, v/v) were used to extract yam. Then the antioxidant activity evaluated through ferrous ion chelating activity, ${\beta}$-carotene bleaching method, lipid peroxidation inhibition and nitric oxide (NO) radical scavenging activity. 70% Methanol extract showed the highest ferrous ion chelating activity and NO radical scavenging activity. And CM extract was the most effective in inhibition of linoleic acid peroxidation evaluated by ${\beta}$-carotene bleaching assay and lipid peroxidation inhibition assay. Based on the results obtained, yam is a potential active ingredient that could be applied in antioxidation as well as bio-health functional food to take a good part in prevention of human diseases and aging.

계면활성제가 돌연변이원성 물질의 활성에 미치는 영향 (Effect of Synthetic Surfactants on the Activity of Mutagens)

  • 최윤호;정용
    • Environmental Analysis Health and Toxicology
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    • 제7권3_4호
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    • pp.69-79
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    • 1992
  • Recently, concerns of water pollution and health risks caused by synthetic detergents have emerged, as the use of various detergents has increased It has been suggested that some surfactants are cocarcinogens. The surfactants tested were linear alkylbenzene sulfonate, sodium lauryl sulfate, polyoxyethylene sorbitan monooleat (tween 80), and the mutagens were 1-nitropyrene, N -methyl- N'-nitro-N -nitrosoguanidine, benzo (a) pyrene, and aflatoxin B$_1$. This study was undertaken to investigate the effects of surfactants on the activity of mytagens using the Ames mutagenic assay with Salmonella typhimurium TA98, TA100. The results were summarized as follows: 1. The surfactants have no mutagenic activity of themselves. 2. Higher doses of surfactants than 100 $\mu\textrm{g}$/plate reduced the number of revertants. It is assumed that the reduction would inhibited cell growth. 3. When the comutagenic ratio is defined as the ratio between mutagenic activity itself and the activity with mutagen and surfactant (drinking water quality standard), LAS showed the comutagenic ratio 0.86-1.17 and SLS 0.74-1.10 as well. According to the comparisons, it could not be recognised for the comutagenicity of drinking water quality standard of surfactant. 4. As increasing the amount of mutagens, the designated amount of surfactant did not affected the mutagen's activity statistically. From the above result, syunthetic surfactants do not present mutagenicity and comutagenicity in the microbial assay.

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