• Title/Summary/Keyword: MTT assay

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Anti-inflammatory Effect of Biotin and Plant extracts

  • Y. J. Joo;S. W. Jung;Kim, B. R.;Kim, I. Y.;Lee, J. D.;H. C. Ryoo;Lee, S. H.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.601-610
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    • 2003
  • Biotin is a water-soluble vitamin used as a skin conditioning agent and promotes the formation of intercellular lipid layers through increased lipid synthesis, which improves the skin's natural barrier function. The anti-inflammatory effects of biotin have been investigated using in vitro assay models, such as MTT assay, measurements of concentrations of nitric oxide(NO), prostaglandin E2(PGE$_2$), and inhibition rate of 5-lipoxygenase(5-LOX). In comparison with biotin, other plant extracts were tested at the same time which were kudzu vine extract, sage extract, paeonia extract, and dipotassium glycyrrhetinate. Nitric oxide is a signal molecule with functions such as neurotransmission, local vascular relaxation, and anti-inflammation in many physiological and pathological processes. NO can cause apoptosis and necrosis of target cells such as keratinocytes and is generated from L-arginine by nitric oxide synthase (NOS). Prostanoids, including prostaglandins and thromboxanes, are generated by the phospholipase $A_2$/cyclooxygenase(COX) pathway, and leukotrienes are generated by the 5-lipoxygenase pathway from arachidonic acid. Prostaglandin E2 recently have been shown to be beneficial in the resolution of tissue injury and inflammation, also has been implicated as an immunosuppressive agent and plasma levels of PGE$_2$ are elevated in patients sustaining thermal injury. Lipoxygenase metabolites from arachidonic acid have been implicated in inflammation, anti-inflammatory activity of the raw materials was evaluated in vitro by the offered inhibition of lipoxygenase.

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Ginsenoside Rb3 ameliorates podocyte injury under hyperlipidemic conditions via PPARδ- or SIRT6-mediated suppression of inflammation and oxidative stress

  • Heeseung Oh;Wonjun Cho;Seung Yeon Park;A.M. Abd El-Aty;Ji Hoon Jeong;Tae Woo Jung
    • Journal of Ginseng Research
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    • v.47 no.3
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    • pp.400-407
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    • 2023
  • Background: Rb3 is a ginsenoside with anti-inflammatory properties in many cell types and has been reported to attenuate inflammation-related metabolic diseases such as insulin resistance, nonalcoholic fatty liver disease, and cardiovascular disease. However, the effect of Rb3 on podocyte apoptosis under hyperlipidemic conditions, which contributes to the development of obesity-mediated renal disease, remains unclear. In the current study, we aimed to investigate the effect of Rb3 on podocyte apoptosis in the presence of palmitate and explore its underlying molecular mechanisms. Methods: Human podocytes (CIHP-1 cells) were exposed to Rb3 in the presence of palmitate as a model of hyperlipidemia. Cell viability was assessed by MTT assay. The effects of Rb3 on the expression of various proteins were analyzed by Western blotting. Apoptosis levels were determined by MTT assay, caspase 3 activity assay, and cleaved caspase 3 expression. Results: We found that Rb3 treatment alleviated the impairment of cell viability and increased caspase 3 activity as well as inflammatory markers in palmitate-treated podocytes. Treatment with Rb3 dosedependently increased PPARδ and SIRT6 expression. Knockdown of PPARδ or SIRT6 reduced the effects of Rb3 on apoptosis as well as inflammation and oxidative stress in cultured podocytes. Conclusions: The current results suggest that Rb3 alleviates inflammation and oxidative stress via PPARδ-or SIRT6-mediated signaling, thereby attenuating apoptosis in podocytes in the presence of palmitate. The present study provides Rb3 as an effective strategy for treating obesity-mediated renal injury.

Diosgenin Inhibits hTERT Gene Expression in the A549 Lung Cancer Cell Line

  • Mohammad, Rahmati Yamchi;Somayyeh, Ghareghomi;Gholamreza, Haddadchi;Majid, Mobasseri;Yousef, Rasmi
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6945-6948
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    • 2013
  • Background: Diosgenin, a steroidal saponin from a therapeutic herb, fenugreek (Trigonellafoenum-graceum L.), has been recognized to have anticancer properties. Telomerase activity is not detected in typical healthy cells, while in cancer cell telomerase expression is reactivated, therefore providing a promising cancer therapeutic target. Materials and Methods: We studied the inhibitory effect of diosgenin on human telomerase reverse transcriptase gene (hTERT) expression which is critical for telomerase activity. MTT- assays and qRT-PCR analysis were conducted to assess cytotoxicity and hTERT gene expression inhibition effects, respectively. Results: MTT results showed that $IC_{50}$ values for 24, 48 and 72h after treatment were 47, 44 and $43{\mu}M$, respectively. Culturing cells with diosgenin treatment caused down-regulation of hTERT expression. Discussion: These results show that diosgenin inhibits telomerase activity by down-regulation of hTERT gene expression in the A549 lung cancer cell line.

Expression of recombinant protein from Oenanthe javanica DC. leaf tissues as a biofactory (Biofactory로서의 미나리 엽조직을 이용한 재조합단백질 발현)

  • Shin, Dong-Il;Park, Hee-Sung
    • KSBB Journal
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    • v.23 no.6
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    • pp.554-556
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    • 2008
  • Fresh Oenanthe javanica DC. leaves still attached to stem architecture were immersed in NaOH solution for 3 min before agroinfiltration and co-cultivation. MTT assay revealed that NaOH solution containing up to 0.7% was still safe for the leaf viability. Fluorometric GUS enzyme analysis showed that 0.5% NaOH-treated leaf tissues were efficiently transformed by vacuum infiltration for 20 min with Agrobacterium cells at a density of $OD_{600}=0.5$ to 1.0. These conditions worked well for the expression of HBsAg, which was confirmed by western blotting and ELISA.

Effect of Retrovirus Mediated TNF-$\alpha$ Gene Transfer to Tumor Necrosis Factor(TNF) Sensitive Tumor Cell Lines on Sensitivity to TNF (Retroviral Vector를 이용한 TNF-$\alpha$ 유전자의 이입이 암세포의 종양괴사인자(TNF) 감수성에 미치는 효과)

  • Oh, Yeon-Mok;Park, Kyeo-Yeong;Jung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Goo;Sim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.2
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    • pp.87-96
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    • 1994
  • Background : Since tumor necrosis factor was discovered in 1975, TNF has been well known about its cytotoxic effect on tumor cells in vivo and in vitro. According to the recent improvement of molecular biological techinques, it is possible that exogenous TNF gene is transferred to tumor cells and is expressed in theirs. By virtue of TNF gene transfer, we have expected that TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells in vivo without systemic side effect. The expected mechanisms in which antitumor effects of TNF expressed in TNF-gene-transferred tumor cells are working would be as followings. In the first mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells around(like homicide). In the second mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill themselves(like suicide). In the third mechanism, TNF expressed in TNF-gene-transferred tumor cells would recruit immune effector cells and kill tumor cells indirectly. In the last mechanism, TNF expressed in TNF-gene-transferred tumor cells would augment cytokine such as interferon-$\gamma$ to kill tumor cells. Among these four mechanisms of antitumor effect, only the second mechanism has not been established yet. Therefore, to elucidate the second mechanism, We performed this study. Method : We transferred TNF-$\alpha$ gene to NCI-H2058, a human mesothelioma cell line and WEHI164, a murine fibrosarcoma cell line by using retroviral vector(pLT12SNTNF). And, We determined by using MTT assay whether TNF expressed in TNF-gene-transferred tumor cell lines would kill themselves like suicide or not. Then, if TNF-gene-transferred tumor cell lines would not suicide themselves, I would know more about the TNF sensitivity of TNF-gene-transferred tumor cell lines to exogenous TNF also by MTT assay. Result : NCI-H2058 and WEHI164 which were sensitive to TNF, became far less sensitive to endogenous and exogenous TNF after being transferred TNF-$\alpha$ gene to. Conclusion : TNF-gene-transfer to NCI-H2058 and WEHI164 gave them resistance to TNF.

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The comparative study on Cell Cytotoxicity of H2O2 and Grapefruit Seed extract (콘택트렌즈 보존제 H2O2와 자몽씨 추출물의 세포 독성 비교 연구)

  • Kim, In-Suk;Yoo, Geun-Chang
    • Journal of Korean Ophthalmic Optics Society
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    • v.9 no.1
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    • pp.173-180
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    • 2004
  • This study aims to investigate the effects of hydrogen peroxide and grapefruit seed extract used as a chemical and natural disinfectants on human conjunctival cells in vitro. The main component of grapefruit seed extract is a narigin. It is one of the flavonoid types in citrus fruits and f1avonoids are widely recognized as naturally occurring(삭제) antioxidants. Cytotoxicity was determined by mitochondrial activity(MTT assay) and DNA damage was analyzed by measuring Comet assay. In LDH assay, 5% of grapefruits seed extract has been observed as a material is giving recovery effect of damaged cultured conjuctival cells by hydrogen peroxide. And also, each of concentrations has been treated simultaneously with same amounts and cytotoxicity of hydrogen peroxide and grapefruit seed extract have been estimated by LDH leakage assay after 24 hours. In conclusion, H2O2-induced cytotoxicity, apoptosis were Significantly prevented by grapefruit seed extract. It is a main component of bioflavonoids that we can simply take it as food. The present results suggest that grapefruit seed extract is a useful disinfectanct having antioxidant and antiapoptopic activity as a natural product.

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Functional Cosmetic Effect of Porcine Placeta (Porcine Placenta의 기능성 화장품소재 특성)

  • Kim, Bo Young;Kim, Tagon;Kang, Whan Yul;Baek, Hyun;Cheon, Hae Young;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.48 no.3
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    • pp.327-331
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    • 2010
  • Porcine placenta was treated with alkali, acid and enzyme treatment to obtain extracts. Heavy metal contents such as Pb, As, and Hg were low enough to satisfy cosmetic agent standard. As a result of safety test(MTT assay), porcine placenta extracts showed over 80% of cell viability at $50{\mu}g/ml$, and cell toxicity was relatively lower. From antioxidation test using DPPH free radical scavenging assay, antioxidation effect was highest as 63% at $50{\mu}g/ml$ when porcine placenta was treated with alkali in pH 9. From whitening effect test using tyrosinase inhibition assay, tyrosinase inhibition effect was 30% at $50{\mu}g/ml$ concentration in alkali treated procine placenta, however, the efficiency was lower compared with arbutin or vitamin C. In anti-wrinkle effect test from elastase inhibition assay, elastase inhibition effects were 20~30% at $50{\mu}g/ml$ for 5 kinds of porcine placenta treatments, which was superior to standard, and especially, protease treated extracts showed best results. Skin formulation including 1% porcine placenta was made and the formulation was very stable for temperature and storage period. From this research, porcine placenta extract showed high potential for anti-wrinkle functional cosmetic agent.

The Antioxidant and Anti-aging Effects of Treatment with Schisandra chinensis Seeds Fractions in UVB-irradiated Human HaCaT Cells (자외선 B에 유도된 사람유래 HaCaT cells에서의 오미자 종자 분획물의 항산화 및 항노화 효과)

  • Choi, Eun-Young;Sohn, Ho-Yong;Lee, Jin-Tae
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1071-1079
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    • 2019
  • In this study, we investigated the antioxidant and anti-aging activities of Schisandra chinensis seed fractions by adding them to UVB-irradiated HaCaT cells and analyzing the suppression of matrix metalloproteinases (MMPs). An electron-donating assay, ABTS radical scavenging assay, and hydrogen peroxide scavenging assay showed that the ethyl acetate fraction of S. chinensis seed (SCEAf) has scavenging activities. A collagenase inhibition activity assay showed that SCEAf had inhibitory effects of over 92.3% at $500{\mu}g/ml$ concentration. An MTT assay was performed to determine the cytotoxicity of SCEAf in HaCaT cells and the results showed that SCEAf increased cellular viability in a concentration-dependent manner. In addition, SCEAf was found to increase the viability of cells irradiatged by UVB $50mJ/cm^2$. To examine the anti-aging effects of SCEAf on HaCaT cells irradiated with UVB $50mJ/cm^2$, the expression of MMP-1 and -3 was analyzed by Western blotting and reverse-transcription polymerase chain reaction. The results showed that MMP-1 and -3 proteins and mRNA levels were downregulated in a concentration-dependent manner in response to SCEAf. These results suggest that SCEAf can prevent aging and alleviate aging symptoms by inhibiting collagenase activity in skin keratinocytes damaged by UVB. Therefore, S. chinensis seeds may have the potential for use as functional ingredients with anti-aging effects in the cosmetic and food industries.

Anticancer Effect of Extracts from the Marine and Salted Fish Products. (수산물 및 수산 발효식품의 암세포 억제효과)

  • 임현수;김수현;유은정;강동수;최명락;송상호
    • Journal of Life Science
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    • v.11 no.1
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    • pp.48-53
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    • 2001
  • This study was performed to observe the cytotoxic effect of the various salted fish extracts against cancer cell line, human hepatocellular carcinoma (HepG2) using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) method. Urechis unicinctus was the strongest cytotoxic effects among any other traditional salted fish products. The growth inhibition ratio of Urechis unicinctus hot-water extracts was 94.5% at the concentration of 1000$\mu\textrm{g}$/$m\ell$. On the other hand, in case of salted fish methanol extracts, salt-fermented shad gizzard was showed the strongest cytotoxic effects. The growth inhibition ratio of salt-fermented shad gizzard methanol extracts was investigated 90% at the concentration of 1000$\mu\textrm{g}$/.$m\ell$.

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Effects of Myrrha Water Extract on Rat Myocardial Cells in Cultures (몰약 전탕액이 배양 심근세포에 미치는 영향)

  • 권강범;조현익;김구환;김상범;이호섭;황우준;박승택;류도곤
    • The Journal of Korean Medicine
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    • v.21 no.2
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    • pp.79-86
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    • 2000
  • Objectives and Methods : In order to elucidate toxic mechanism of myocardial damage and protective effect of myrrha water extract against cytotoxic effect of xanthine oxidase/hypoxanthine(XO/HX), cardioprotective effect of myrrha water extract was examined by MTT assay, LDH (Lactate Dehydrogenase) activity and heart beating rate after cultured myocardial cells derived from neonatal mouse were treated with various concentration of XO/HX, a free radical. Results : XO/HX induced a decrease of cell viability, an increase in the amount of LDH, and a decrease of heart beating rate on cultured myocardial cells in a dose-dependent manner. In cardioprotective effect of myrrha water extract, it showed a decrease in the amount of LDH and an increase of heart beating rate on cultured myocardial cells damaged by XO/HX. Conclusions : From the above results, it is suggested that XO/HX showed toxic effect in cultured myocardial cells derived from neonatal mouse and that myrrha water extract is very effective in the prevention of XO/HX-induced cardiotoxicity.

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