• Title/Summary/Keyword: Inhibition ELISA

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Inhibitory effect of broccoli leaf extract on PGE2 production by NF-κB inhibition (NF-κB 저해를 통한 브로콜리 잎 추출물의 PGE2 저해효과)

  • Park, Sook Jahr;An, Iseul;Noh, Gyu Pyo;Yoo, Byung Hyuk;Lee, Jong Rok
    • The Korea Journal of Herbology
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    • v.34 no.6
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    • pp.117-124
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    • 2019
  • Objective : Broccoli is edible green plant that has a wide variety of health benefits including cancer prevention and cholesterol reduction. However, leaves of broccoli are not eaten and are mostly left as waste. This study was conducted to evaluate the effects of the broccoli leaf extract (BLE) on prostaglandin E2 (PGE2) production related to nuclear factor kappa B (NF-κB) signaling in lipopolysaccharide (LPS)-activated macrophages. Methods : BLE was prepared by extracting dried leaf with ethanol. Cell viability was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. PGE2 and inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA). Expression level of each protein was monitored by Western blot analysis. Results : In LPS-activated Raw264.7 cells, PGE2 release into culture medium was dramatically enhanced compared to control cells. However, increased PGE2 was attenuated dose-dependently by treatment with BLE. Inhibition of PGE2 production by BLE was due to the suppression of cyclooxygenase-2 (COX-2) expression determined by Western blot analysis. BLE also inhibited the production of inflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α). Inhibition at PGE2 and cytokine was mediated from inhibition of nuclear translocation of NF-κB due to the repression of inhibitory kappa B alpha (IκBα) phosphorylation and degradation. Conclusion : This study showed that BLE exerted inhibitory activities against PGE2, which is critical for the initiation and resolution of inflammatory responses, and that inhibition of PGE2 was mediated by suppression of NF-κB signaling. These results suggest that the waste broccoli leaves could be used for controlling inflammation.

Cytotoxicity, Apoptosis Induction and Anti-Metastatic Potential of Oroxylum indicum in Human Breast Cancer Cells

  • Kumar, D.R. Naveen;George, V. Cijo;Suresh, P.K.;Kumar, R. Ashok
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2729-2734
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    • 2012
  • Despite clinical advances in anticancer therapy, there is still a need for novel anticancer metabolites, with higher efficacy and lesser side effects. Oroxylum indicum (L.) Vent. is a small tree of the Bignoniaceae family which is well known for its food and medicinal properties. In present study, the chemopreventive properties of O. indicum hot and cold non-polar extracts (petroleum ether and chloroform) were investigated with MDA-MB-231 (cancer cells) and WRL-68 (non-tumor cells) by XTT assay. All the extracts, and particularly the petroleum ether hot extract (PHO), exhibited significantly (P<0.05) higher cytotoxicity in MDA-MB-231 when compared to WRL-68 cells. PHO was then tested for apoptosis induction in estrogen receptor (ER)-negative (MDA-MB-231) and ER-positive (MCF-7) breast cancer cells by cellular DNA fragmentation ELISA, where it proved more efficient in the MDA-MB-231 cells. Further, when PHO was tested for anti-metastatic potential in a cell migration inhibition assay, it exhibited beneficial effects. Thus non-polar extracts of O. indicum (especially PHO) can effectively target ER-negative breast cancer cells to induce apoptosis, without harming normal cells by cancer-specific cytotoxicity. Hence, it could be considered as an extract with candidate precursors to possibly harness or alleviate ER-negative breast cancer progression even in advanced stages of malignancy.

Survey of foot-and-mouth disease virus structural protein antibody titer in Yeongcheon (영천지역 구제역 바이러스 구조단백질 항체가 조사)

  • Sohn, Jun-Hyung;Hwang, You-Sun;Sohn, Kyu-Hee;Shin, Sung-Ho;Lee, Eun-Mi;Kim, Soon-Tae;Cho, Min-Hee;Yun, Mun-Jo
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.13-17
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    • 2015
  • Three serotypes (O, A and Asia1) of the foot-and-mouth disease (FMD) vaccine were injected into domestic cloven-hoofed animals in korea after the nationwide spread at the end of 2010. The purpose of this study was survey of FMD virus stuructural protein (SP) antibody titer in Yeongcheon by enzyme-linked immunosorbent assay (ELISA). Total 1,324 samples collected from 89 farms were tested. The overall seroprevalence of FMD virus SP antibodies was 58.8% (778/1,324) The seroprevalence of FMD virus SP antibody varied with species. Results in cattle (over 12 month old) and pig (90 to 130 day old) were 58.8% and 44.9% respectively.

Suppressive Effects of Triterpenoids on CINC-1 Induction in Interleukin-1{\betha}-stimulated rat fibroblast NRK-49F cells

  • Ha, Joo-Young;Min, Kyung-Rak;Kang, She-Hoon;Kim, Ju-Sun;Lee, Gyeong-Im;Kang, Sam-Sik;Kim, Youngsoo
    • Archives of Pharmacal Research
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    • v.20 no.3
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    • pp.234-238
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    • 1997
  • CINC-1 is a member of chemokine family with chemotactic and activating properties to neutrophils. CINC-1 induction in IL-$1{\beta}$-stimulated rat fibroblast NRK-49F cells was quantitated by a sensitive ELISA. CINC-1 production was increased up to 135 ng/ml from basal 2-6 ng/ml by stimulation with IL-$1{\beta}$.Steroidal anti-inflammatory drugs including dexamethasone and prednisolone exhibited potent suppressive effects on IL-$1{\beta}$-induced CINC-1 production. Among 39 kinds of natural triterpenoids tested, acacigenin B exhibited the highest suppressive effects with about $10{\mu}M$ to be 50% of inhibition on the CINC-1 induction. The suppressive potency of acacigenin B on IL-$1{\beta}$induced CINC-1 production was about 10-fold less than that of the steroidal anti-inflammatory drugs.

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Effects of Draconis Resina on the Collagenase Activities and the Procollagen Synthesis in Hs68 Human Fibroblasts, and Tyrosinase Activity

  • Kim, Tae Yeon;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.30 no.6
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    • pp.1-6
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    • 2015
  • Objectives : Draconis Resina (DR), the resin of Daemonorops draco Bl., is used to circulate the blood and to stop bleeding. It also has been used to generate flesh including ulceration. The present study investigated the effects of DR extract on collagen metabolism in human fibroblasts and tyrosinase activity in mushroom tyrosinase.Methods : The effect of DR extract on type I procollagen production (collagen type I synthesis) and collagenase (matrix metalloproteinase-1, henceforth referred as MMP-1) activity in human normal fibroblasts cell line. Hs68 cells after ultraviolet B (UVB, 312 nm) irradiation was measured using the enzyme - linked immunosorbent assay (ELISA). The tyrosinase activity was also measured to find out the whitening effects in mushroom tyrosinase by ELISA method.Results : There was no cytotoxicity at DR extract at concentrations of 10 μg/ml, 30 μg/ml, and 100 μg/ml. DR extract significantly inhibited the increase of collagenase activity, whereas it did not show on the reduction of type I procollagen in UVB damaged Hs68 cells. DR extract did not reduce the L - DOPA oxidation. However, it significantly reduced the tyrosinase activity by DR extract at concentraions of 0.1 mg/ml, 1 mg/ml and 10 mg/ml.Conclusions : In conclusion, DR showed the anti-wrinkle and whitening effects via the inhibition of collagenase production and the tyrosinase activity. These results suggest that DR may have potential as an anti-aging ingredient in cosmetic herb markets.

Identification of the IL-1$\beta$ inhibitor in the febrile patient urine by anti-IL-1$\beta$ monoclonal antibody (Anti-IL-1$\beta$ 단일클론 항체를 이용해서 발열환자의 뇨중 IL-1$\beta$ inhibitor의 확인)

  • 남경수;배윤수;남명수;오은숙;박순희;최인성;정태화
    • YAKHAK HOEJI
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    • v.37 no.4
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    • pp.420-426
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    • 1993
  • To effectively purify of IL-1 inhibitor from human febrile urine, we have established monoclonal antibody that reacts with human recombinant interleukin l$\beta$(IL-1$\beta$). The antibody, designated ON-1, was highly specific to IL-1$\beta$ and no cross-reaction with other cytokines(IL-l$\alpha$ and IL-4) was observed. As the results of ELISA inhibition assay and Western blotting method, it was further identified that ON-1 had high binding specificity with IL-1$\beta$. IL-1 receptor binding material from febrile patient urine was effectively purified with affinity column chromatography which conjugated with ON-1. This urinary material inhibited the thymocyte proliferation in a dosedependent manner. IL-l$\beta$ induced thymocyte proliferation activity was inhibited to 67.3% at 6 $\mu\textrm{g}$ of the purified urinary material. The result may suggest that this urinary material the purified urinary material. The result may suggest that this urinary material will have antagonic effect on IL-1 action mechanism and act IL-l$\beta$ inhibitor.

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Serologic monitoring of animal welfare-oriented laying-hen farms in South Korea

  • So, Hyunhee;Jeong, Seolryung;Mo, Jongsuk;Min, Kyungchul;Kim, Jongnyeo;Mo, In-Pil
    • Korean Journal of Veterinary Research
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    • v.58 no.4
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    • pp.193-199
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    • 2018
  • As animal welfare issue becomes important, the European Union bans conventional cages for laying hens from 2012. So the alternative housing systems like floor pens, aviaries or free range systems have been suggested. From 2011 to 2014, we monitored 20 welfare-oriented laying hen farms in South Korea to figure out serological status of major viral diseases. During this period, total 3,219 blood samples were collected from the randomly selected chickens to test and evaluate the hemagglutination inhibition titers for low pathogenic avian influenza, Newcastle disease and egg drop syndrome '76. A total of 2,926 blood samples were tested through enzyme linked immunosorbent assay (ELISA) to assess the serological status of infectious bronchitis (IB). The distribution of ELISA titers for IB was various from almost 0 to 20,000 through the all weeks of age. Also, the antibody coefficient of variation for most of the diseases in this study was higher than those of typical cage layers. As this study was the first surveillance for major avian viral diseases of the animal welfare-oriented farms in South Korea, the results obtained from this study will help to determine what information and resources are needed to maintain better biosecurity and to improve the health and welfare of laying hen flocks.

Interleukin-5 Inhibition Assay of the Oriental Materia Medica Treatment by Sandwich ELISA on Mouse Splenocytes (ELISA를 이용한 IL-5 분비조절 한약물 Screening)

  • Park, Ki-Bok;Jung, Sung-Ki;Jung, Hee-Jae
    • The Journal of Internal Korean Medicine
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    • v.30 no.3
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    • pp.582-593
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    • 2009
  • Background and Objective : Allergy is defined as an altered reactivity to an antigen, which can result in pathologic reactions upon subsequent exposure to that particular antigen. This study was to evaluate the effect of oriental materia medica on IL-5 secretion in the mouse spleen cell. Methods : We used the splenocytes of mouse 8 weeks after its birth, and then cultivated those into the 2 experimental groups and a control group for 48 hours. The culture media of the experimental groups were made of $1{\mu}g/ml$, $10{\mu}g/ml$ oriental materia medica, representative. And the culture media of control group was given no oriental materia medica. Then, we assayed the quantity of cytokine-expression by the sandwich ELISA. The quantities of cytokine-expression of the experimental groups were compared with that of the control group, which was standardized. These methods were used for all of the oriental materia medica treated. Results : Some oriental materia medica inhibit the secretion of IL-5 in both $1{\mu}g/ml$ and $10{\mu}g/ml$ culture media. These were Acori Rhizoma, Luffae Fasciculus Vascularis, Amomi Rotundi Fructus, Schisandrae Fructus, Biotae Semen, Clematis armandii, Dioscoreae Sativa Rhizoma, Coicis Semen, Sophorae Flos, Oroxyli Semen, Aurantii Semen, Pini Nodi Lignum. Conclusion : This study indicates that some oriental materia medica inhibit the secretion of IL-5 and are beneficial for allergic disease.

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PCR technique for detection of toxigenic Pasteurella multocida in mixed bacterial cultures from pigs (Polymerase chain reaction을 이용한 독소생산성 Pasteurella multocida의 검출)

  • Chi, Yongzhe;Lee, Dong-seok;Han, Jeong-hee;Han, Kyung-soo;Hahn, Tae-wook
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.56-62
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    • 2000
  • Pasteurella multocida is kind of commensal bacteria in the upper respiratory tract of pigs. It is classified toxigenic and nontoxigenic strains based on the production of dermonecrotic toxin. Toxigenic strain is most associated with atrophic rhinitis which brings great economical loss in swine industry. However, toxigenic and nontoxigenic strains do not differ by diagnostic biochemical reaction or morphology. One of recently developed techniques, PCR detects the toxigenic P multocida. Amplification of an 846-nucleotide fragment of toxA gene was developed. The fragment amplified by PCR was detected in P multocida type D not type A. The PCR amplification was as sensitive as it could detect 1 pg of P multocida DNA. We compared the result of the PCR with the enzyme linked immunosorbent assay (ELISA) in a test for 40 swine nasal swabs. All of these isolates were toxin negative based on the ELISA while 2 isolates were detected in the PCR technique. in addition to accuracy, as required for rapid detection from contaminated nasal swabs, toxigenic P multocida was recovered efficiently from contaminated culture without inhibition of the PCR. The results show that the PCR detection of toxigenic P multocida directly form nasal swabs are feasible.

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A Study on Antibacterial Activity and Seroprevalence of Ornithobacterium rhinotracheale Isolated from the Domestic Chickens (국내 사육 닭에서 분리된 Ornithobacterium rhinotracheale (OR)균의 약제 감수성 및 항체보유율에 대한 연구)

  • 전우진;권용국;윤여성;김재홍
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.161-165
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    • 2003
  • Ornithobacterium rhinotracheale (OR) is a recently described gram-negative rod-shaped bacterium associated with respiratory tract infection in poultry. In order to investigate current occurrence of OR infection and to evaluate antibiotic susceptibility, the prevalence of OR antibody in domestic chickens were examined and the minimal inhibitory concentrations(MICs) of 8 antibiotics for 11 OR isolates was determined. All isolates tested were mostly susceptible to three antibiotics, ampicillin (MICs ranging from 0.38 ${\mu}g$/ml to 2 ${\mu}g$/ml), tetracycline (MICs 0.094~3 ${\mu}g$/ml) and doxycycline (MICs 0.047~4 ${\mu}g$/ml) but resistant to genatmicin. Ciprofloxacin, norfloxacin, enrofloxacin, and ofloxacin gave most isolates inhibition only in case of a higher concentration (MICs ranged in most cases from 3 ${\mu}g$/ml to 48 ${\mu}g$/ml). Out of 188 chicken flocks including broilers, broiler breeders, and layers, seropositive flock to OR were detected in 5 broilers (4%), 17 broiler breeders (50%), and 16 layers (55.2%), using commercial OR enzyme-linked immunosorbent assay(ELISA) kits. It suggested that OR infection was widespreaded in poultry farms in Korea.