• Title/Summary/Keyword: Activity assay

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삼백초 물 추출물과 유기용매 분획물의 항산화, 항염증 및 PMA에 의해 유도된 MMP-2 및 MMP-9활성 침윤 억제 효과 (Effect of Anti-oxidant, Anti-inflammatory and Anti-invasive of PMA-induced Matrix Metalloproteinase (MMP-2) and MMP-9 Activities of Water Extract and Solvent Fractions of Saururus Chinensis)

  • 김준호;김은정
    • 생명과학회지
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    • 제26권5호
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    • pp.584-591
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    • 2016
  • 삼백초는 항산화 활성을 갖는 플라보노이드 화합물을 포함하고 있는 여러해살이 식물로 알려져 있다. 본 연구에서는 삼백초 물 추출물과 유기용매 분획물의 항산화 활성, 항염증 활성 및 PMA에 의해 유도된 MMP-2 및 MMP-9활성 억제 효과를 조사하였다. 시료들은 삼백초 물 추출을 헥산(hexane), 클로로포름(CHCl3), 에틸 아세테이트(Ethyl acetate), 부탄올(butanol) 및 물(water)과 같은 용매로 분획화하여 사용하였고, 물 분획물의 수득율이 9.25%로 가장 높았다. 항산화 활성은 DPPH assay, 세포 생존율 측정은 MTS assay, 항염증 활성은 마우스 대식세포 Raw 264.7세포에서NO 생성 그리고 MMP-2 및 MMP-9의 mRNA 발현 및 단백질 활성 억제는 인간구강편평세포암종 YD-10B 세포에서 RT-PCR과 zymography방법을 통해 측정하였다. 본 연구의 결과에 의하면 MMP-2/-9 활성은 PMA에 의해 YD-10B세포에서 증가하였다. PMA 처리된 YD-10B 세포에서, 에틸 아세테이트 분획물이 증가된 MMP-2/-9의 mRNA 발현 및 단백질 활성을 유의하게 억제하였다. 그리고 항산화 활성도 에틸아세트 분획물이 73.38%의 가장 높게 나타났다. 또한 세포 독성을 나타내지 않는 농도에서, 에틸아세트 분획물이 Raw 264.7세포에서 농도 의존적으로 유의하게 항염증 활성을 보였다. 그러므로 본 연구에서는 삼백초 물 추출의 에틸아세트 분획물이 구강암의 암 침윤을 억제할 수 있는 효과적인 암 예방 및 치료를 위한 항암제로서의 가능성을 암시하고 있다.

α-Asarone이 항산화 활성 및 기질금속단백질 분해효소 활성 조절에 미치는 영향 (α-Asarone Modulates Activity of Matrix Metalloproteinase as well as Antioxidant Activity)

  • 박혜정;김문무
    • 생명과학회지
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    • 제25권9호
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    • pp.1000-1006
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    • 2015
  • α-Asarone은 동양의 전통적인 약재로 잘 알려진 석창포(Acorus gramineus)의 주된 성분이다. 석창포는 항위궤양, 항알러지, 히스타민 방출 억제 그리고 항산화 효과와 같이 다양한 효과를 나타내는 것으로 잘 알려져 있다. 그러나 석창포 역할에 대한 기전연구는 아직 부족한 실정이다. 본 연구에서는, HT1080 세포주에서 α-asarone의 항산화 효과뿐만 아니라 matrix metalloproteinase에 대한 효과를 조사하였다. 가장 먼저 α-asarone의 세포 생존에 대한 효과를 조사하기 위해 MTT assay를 이용하여 16 μM이하에서 세포독성이 없음을 나타내었다. α-asarone이 환원력과 fenton reaction에 의해 유도된 DNA 산화로부터 보호효과를 나타내는 것을 확인하였다. 더욱이, α- asarone은 collagenase 활성을 증가시키고 phorbol 12-myristate 13-acetate (PMA)로 자극된 MMP-2 및 MMP-9의 활성을 증가시켰다. 한편 phenazine methosulfate (PMS) 로 자극된 경우 MMP-9의 활성은 α-asarone의 존재하에서 증가되었으나 MMP-2 활성에는 변화가 없었다. 그러므로 우리의 연구결과는 α-asarone이 산화적 스트레스 및 MMPs와 관련된 병리학적 질환의 예방 및 치료제로 개발이 기대된다고 제안한다.

Post HCV Infection Due to MX Gene Stimulation Produced Post Treatment with Imported and Locally Produced Egyptian Biosimilar IFN

  • Mohamed, Shereen H;Mahmoud, Nora F;Mohamed, Aly F;Kotb, Nahla S
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권14호
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    • pp.5635-5641
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    • 2015
  • Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.

홍삼 추출물이 두부의 저장성과 항산화 활성에 미치는 영향 (Effect of Red Ginseng Extract on Storage and Antioxidant Activity of Tofu)

  • 이정숙;김교남;장해동
    • 한국식품영양과학회지
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    • 제37권11호
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    • pp.1497-1506
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    • 2008
  • 홍삼 추출물을 첨가한 두부를 제조함에 있어 홍삼 추출물 첨가량이 두부의 품질과 저장성 그리고 항산화 활성에 미치는 영향을 조사하였다. 건조 대두를 기준으로 홍삼 추출물을 $0.5{\sim}2%$ 첨가하여 제조한 두부와 첨가하지 않은 두부의 물성, 색도, 관능의 차이를 비교하였다. 두부의 texture는 견고성과 탄력성에서만 홍삼 추출물의 첨가량에 따라 다소 증가하는 경향으로 홍삼 추출물을 2% 첨가한 두부에서 유의적 차이가 나타났고, 두부의 색도는 L, a, b값 모두 유의적으로 증가하였다. 관능검사 결과 color에 대한 기호도가 떨어지는 경향을 보여 주었으나 홍삼 특유의 맛이 유의적으로 감지되면서 전체적인 기호도는 통계상 유의적 차이가 보이지 않아 홍삼 추출물의 첨가가 소비자의 기호에 특별한 거부감을 일으키지 않는 것으로 나타났다. 두부를 $15^{\circ}C$에서 보관하면서 pH, 탁도 그리고 총 균수의 변화를 측정한 결과 저장 초기에는 변화가 없다가 pH와 총 균수는 저장 4일부터, 탁도는 저장 6일부터 변화하는 것으로 나타나, 이는 홍삼 추출물이 첨가된 두부가 홍삼 추출물이 첨가되지 않은 두부보다 미생물 생육이 억제되어 두부의 저장성이 향상되었음을 의미한다. ORAC assay를 이용한 두부의 항산화 활성은 첨가된 홍삼 추출물의 농도에 비례하여 antioxidant 활성이 유의적으로 증가하는 경향을 나타내었으며, ORACOH. assay에서 는 홍삼 추출물을 첨가한 두부 추출물이 홍삼 추출물의 농도에 비례하여 pro-oxidant 활성을 보여 주었다.

화장품 소재로서 빈랑자의 항산화 및 미백활성에 관한 효과 (A Study of the Antioxidant Activities and Whitening Activities of Areca semen Extracts as Cosmetic Ingredient)

  • 강희철;차미연;김재영
    • 대한화장품학회지
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    • 제41권3호
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    • pp.269-277
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    • 2015
  • 항산화 활성과 미백활성을 갖는 새로운 화장품 소재를 찾기 위하여 한방 소재로 사용되는 빈랑자[Areca semen (A. semen)]를 유기용매 추출을 통하여 활성분획을 분리하였다. 추출된 활성 분획을 이용하여 항산화 활성, 피부 상재균에 대한 항균 활성, 세포독성 및 세포 내 멜라닌 합성 저해능 및 멜라닌 합성에 관여하는 유전자의 저해 활성을 확인함으로써 미백 화장품 소재로서의 가능성을 확인하였다. 빈랑자는 에틸아세테이트 용매의 분획을 동결 건조하여 분말형태로 제조하였으며 이를 이후 실험에 사용하였다. 빈랑자 추출물에서 페놀성 물질은 $301.35{\pm}0.88{\mu}g/mg$으로 측정되었다. 항산화 효과는 DPPH assay와 FRAP assay를 이용하여 측정하였으며, 대조군으로 사용된 ascorbic acid와 비슷하게 측정되었다. 피부상재균에 대한 항균활성은 Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis)에서 모두 $80{\mu}g/mL$의 농도에서 항균활성을 갖는 것으로 나타났으며 Escherichia coli (E. coli)에서는 항균활성을 나타내지 않았다. 미백소재로서의 활성을 확인하여 B16/F1 mouse melanoma cell을 이용하였으며 최대 처리농도인 $80{\mu}g/mL$에서는 독성을 나타내지 않았다. 또한 세포 내 멜라닌 합성 저해능은 $80{\mu}g/mL$에서 $29.44{\pm}0.71%$로 측정되었다. 또한 멜라닌 합성에 관여하는 유전자의 발현량은 mRNA expression assay를 통하여 확인하였으며, 그 결과 tyrosinase와 MITF 유전자는 농도 의존적으로 감소하는 경향을 나타내었다. 따라서 이러한 결과를 통하여 빈랑자의 에틸아세테이트 분획은 항산화와 미백효과를 나타내는 것으로 확인되었으며 화장품 소재로서의 사용 가능성을 확인하였다.

The beneficial effect of ginsenosides extracted by pulsed electric field against hydrogen peroxide-induced oxidative stress in HEK-293 cells

  • Liu, Di;Zhang, Ting;Chen, Zhifei;Wang, Ying;Ma, Shuang;Liu, Jiyun;Liu, Jingbo
    • Journal of Ginseng Research
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    • 제41권2호
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    • pp.169-179
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    • 2017
  • Background: Ginsenosides are the main pharmacological components of Panax ginseng root, which are thought to be primarily responsible for the suppressing effect on oxidative stress. Methods: 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and oxygen radical absorption capacity were applied to evaluate the antioxidant activities of the ginsenosides. Human embryonic kidney 293 (HEK-293) cells were incubated with ginsenosides extracted by pulsed electric field (PEF) and solvent cold soak extraction (SCSE) for 24 h and then the injury was induced by $40{\mu}M$ $H_2O_2$. The cell viability and surface morphology of HEK-293 cells were studied using MTS assay and scanning electron microscopy, respectively. Dichloro-dihydro-fluorescein diacetate fluorescent probe assay was used to measure the level of intracellular reactive oxygen species. The intracellular antioxidant activities of ginsenosides were evaluated by cellular antioxidant activity assay in HepG2 cells. Results: The PEF extracts displayed the higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and stronger oxygen radical absorption capacity (with an oxygen radical absorption capacity value of $14.48{\pm}4.04{\mu}M\;TE\;per\;{\mu}g/mL$). The HEK-293 cell model also suggested that the protective effect of PEF extracts was dose-dependently greater than SCSE extracts. Dichloro-dihydro-fluorescein diacetate assay further proved that PEF extracts are more active (8% higher than SCSE extracts) in reducing intracellular reactive oxygen species accumulation. In addition, scanning electron microscopy images showed that the HEK-293 cells, which were treated with PEF extracts, maintained more intact surface morphology. Cellular antioxidant activity values indicated that ginsenosides extracted by PEF had stronger cellular antioxidant activity than SCSE ginsenosides extracts. Conclusion: The present study demonstrated the antioxidative effect of ginsenosides extracted by PEF in vitro. Furthermore, rather than SCSE, PEF may be more useful as an alternative extraction technique for the extraction of ginsenosides with enhanced antioxidant activity.

Antioxidant Activity of Porcine Skin Gelatin Hydrolyzed by Pepsin and Pancreatin

  • Chang, Oun Ki;Ha, Go Eun;Jeong, Seok-Geun;Seol, Kuk-Hwan;Oh, Mi-Hwa;Kim, Dong Wook;Jang, Aera;Kim, Sae Hun;Park, Beom-Young;Ham, Jun-Sang
    • 한국축산식품학회지
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    • 제33권4호
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    • pp.493-500
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    • 2013
  • Gelatin is a collagen-containing thermohydrolytic substance commonly incorporated in cosmetic and pharmaceutical products. This study investigated the antioxidant activity of gelatin by using different reagents, such as 2,2-azinobis-(3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS), 2,2-di (4-tert-octylphenyl)-1-picrylhydrazyl (DPPH), and oxygen radical absorbance capacity-fluorescein (ORAC-FL) in a porcine gelatin hydrolysate obtained using gastrointestinal enzymes. Electrophoretic analysis of the gelatin hydrolysis products showed extensive degradation by pepsin and pancreatin, resulting in an increase in the peptide concentration (12.1 mg/mL). Antioxidant activity, as measured by ABTS, exhibited the highest values after 48-h incubation with pancreatin treatment after pepsin digestion. Similar effects were observed at 48 h incubation, that is, 61.5% for the DPPH assay and 69.3% for the ABTS assay. However, the gallic acid equivalent (GE) at 48 h was $87.8{\mu}M$, whereas $14.5{\mu}M$ GE was obtained using the ABTS and DPPH assays, indicating about sixfold increase. In the ORACFL assay, antioxidant activity corresponding to $45.7{\mu}M$ of trolox equivalent was found in the gelatin hydrolysate after 24 h hydrolysis with pancreatin treatment after pepsin digestion, whereas this activity decreased at 48 h. These antioxidant assay results showed that digestion of gelatin by gastrointestinal enzymes prevents oxidative damage.

Antimicrobial, Antioxidant and Cytotoxic Activities of Dendropanax morbifera Léveille extract for mouthwash and denture cleaning solution

  • Kim, Ryeo-Woon;Lee, Sook-Young;Kim, Su-Gwan;Heo, Yu-Ri;Son, Mee-Kyoung
    • The Journal of Advanced Prosthodontics
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    • 제8권3호
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    • pp.172-180
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    • 2016
  • PURPOSE. The purpose of this study was to analyze the antimicrobial, antioxidant activity and cytotoxicity of Dendropanax morbifera $L{\acute{e}}veille$ extract for assessing whether Dendropanax morbifera $L{\acute{e}}veille$ can be used for the development of natural mouthwash and denture cleaning solution. MATERIALS AND METHODS. The extract was obtained from branches of Dendropanax morbifera $L{\acute{e}}veille$. The solvent fractions were acquired by fractionating Dendropanax morbifera $L{\acute{e}}veille$ extract using n-hexane, ethyl acetate, chloroform and butanol solvent. Paper disc test was used to evaluate the antimicrobial and antifungal activity of Dendropanax morbifera $L{\acute{e}}veille$ extract and solvent fractions against Streptococcus mutans and Candida albicans. The analysis of antioxidant activity was carried out through DPPH radical scavenging assay. The cytotoxicity of Dendropanax morbifera $L{\acute{e}}veille$ extract was analyzed through MTT assay using normal human oral keratinocytes. RESULTS. Dendropanax morbifera $L{\acute{e}}veille$ extract showed antimicrobial activity against Streptococcus mutans and especially Candida albicans. The solvent fractions of Dendropanax morbifera $L{\acute{e}}veille$ showed strong antimicrobial activity against Streptococcus mutans and Candida albicans in n-hexane and butanol solvent fraction, respectively. Dendropanax morbifera $L{\acute{e}}veille$ extract also showed outstanding antioxidant activity. Butanol, ethyl acetate, and chloroform solvent fraction of Dendropanax morbifera $L{\acute{e}}veille$ tended to have increased antioxidant activity as the concentration increased. Dendropanax morbifera $L{\acute{e}}veille$ extract showed high cell survival rate in cytotoxicity test. CONCLUSION. Dendropanax morbifera $L{\acute{e}}veille$ extract turned out to have antimicrobial, antioxidant activity and cytophilicity. Based on these results, it is expected that Dendropanax morbifera $L{\acute{e}}veille$ is applicable as an ingredient for natural mouthwash and denture cleanser.