• Title/Summary/Keyword: Inhibition assay of ELISA.

Search Result 171, Processing Time 0.021 seconds

Anti-inflammatory Effects of the Water Extract of Phyllostachys Folium via NF-κB Inhibition (죽엽 열수추출물의 염증억제 효과)

  • Son, Jin Won;Park, Sang Mi;Jung, Ji Yun;Hwangbo, Min;Cho, Il Je;Jung, Tae Young;Park, Chung A;Kim, Sang Chan;Jee, Seon Young
    • Herbal Formula Science
    • /
    • v.24 no.4
    • /
    • pp.259-269
    • /
    • 2016
  • Objectives : Phyllostachys Folium is leaves of Phyllostachys nigra var. henesis $S_{TAPF}$. In the East Asian traditional medicine, the herb has been used to treat nasal bleeding, dysuria, epilepsy and etc. The present study was conducted to evaluate the anti-inflammatory effects of the Phyllostachys Folium water extracts (PFE) in vitro and in vivo model. Methods : Cell viability was measured by MTT assay after the treatment of PFE and NO production was monitored by measuring the nitrite content in culture medium. iNOS, COX-2, $I{\kappa}B$, $p-I{\kappa}B{\alpha}$ amd $NF{\kappa}B$ were detected by immunoblot analysis, and levels of cytokine were analyzed by sandwich ELISA kit. Anti-edema effect of PFE was determined in the carrageenan-induced paw edema model in rats. Results : LPS increased NO and cytokines levels compared with control, these increases were attenuated by PFE. In addition, LPS-induced pro-inflammatory proteins such as iNOS, COX-2 were down regulated by PFE. These anti-inflammatory effect of PFE results from inhibition of phosphorylation of $I{\kappa}B$ and translocation of $NF-{\kappa}B$. Conclusion : These results show that PFE has some anti-inflammatory effects which might play a role in gram-negative bacterial infection inflammation and $NF{\kappa}B$ activated diseases.

Inhibitory Effect of Chitosan and Phosphate Cross-linked Chitosan against Cucumber Mosaic Virus and Pepper Mild Mottle Virus

  • Gangireddygari, Venkata Subba Reddy;Chung, Bong Nam;Cho, In-Sook;Yoon, Ju-Yeon
    • The Plant Pathology Journal
    • /
    • v.37 no.6
    • /
    • pp.632-640
    • /
    • 2021
  • Cucumber mosaic virus (CMV) and Pepper mild mottle virus (PMMoV) causes severe economic loss in crop productivity of both agriculture and horticulture crops in Korea. The previous surveys showed that naturally available biopolymer material - chitosan (CS), which is from shrimp cells, reduced CMV accumulation on pepper. To improve the antiviral activity of CS, it was synthesized to form phosphate cross-linked chitosan (PCS) and compared with the original CS. Initially, the activity of CS and PCS (0.01%, 0.05%, and 0.1% concentration) compound against PMMoV infection and replication was tested using a half-leaf assay on Nicotiana glutinosa leaves. The total number of local lesions represented on a leaf of N. glutinosa were counted and analyzed with phosphate buffer treated leaves as a negative control. The leaves treated with a 0.1% concentration of CS or PCS compounds exhibited an inhibition effect by 40-75% compared with the control leaves. The same treatment significantly reduced about 40% CMV accumulation measured by double antibody sandwich enzyme-linked immunosorbent assay and increased the relative expression levels of the NPR1, PR-1, cysteine protease inhibitor gene, LOX, PAL, SRC2, CRF3 and ERF4 genes analyzed by quantitative reverse transcriptase-polymerase chain reaction, in chili pepper plants.

Protective Role of Fucoidan in Cerebral Ischemia-Reperfusion Injury through Inhibition of MAPK Signaling Pathway

  • Che, Nan;Ma, Yijie;Xin, Yinhu
    • Biomolecules & Therapeutics
    • /
    • v.25 no.3
    • /
    • pp.272-278
    • /
    • 2017
  • Fucoidan has been reported to exhibit various beneficial activities ranging from to antivirus and anticancer properties. However, little information is available about the effects of fucoidan on cerebral ischemia-reperfusion injury (IRI). Our study aimed to explore the effects of fucoidan on cerebral IRI, as well as the underlying mechanisms. Sprague-Dawley (SD) rats were randomly subjected to four groups: Sham, IRI+saline (IRI+S), IRI+80 mg/kg fucoidan (IRI+F80), and IRI+160 mg/kg fucoidan (IRI+F160). Fucoidan (80 mg/kg or 160 mg/kg) was intraperitoneally injected from 7 days before the rats were induced to cerebral IRI model with middle cerebral artery occlusion (MCAO) method. At 24 h after reperfusion, neurological deficits and the total infarct volume were determined. The levels of inflammation-associated cytokines (interleukin (IL)-$1{\beta}$, IL-6, myeloperoxidase (MPO), and tumor necrosis factor (TNF)-${\alpha}$), oxidative stress-related proteins (malondialdehyde (MDA) and superoxide dismutase (SOD)) in the ischemic brain were measured by enzyme-linked immunosorbent assay (ELISA). Besides, the levels of apoptosis-related proteins (p-53, Bax, and B-cell lymphoma (Bcl)-2) and mitogen-activated protein kinase (MAPK) pathway (phosphorylation-extracellular signal-regulated kinase (p-ERK), p-c-Jun N-terminal kinase (JNK), and p-p38) were measured. Results showed that administration of fucoidan significantly reduced the neurological deficits and infarct volume compared to the IRI+S group in a dose-dependent manner. Also, fucoidan statistically decreased the levels of inflammation-associated cytokines, and oxidative stress-related proteins, inhibited apoptosis, and suppressed the MAPK pathway. So, Fucoidan plays a protective role in cerebral IRI might be by inhibition of MAPK pathway.

Decreased Expression of TRPV4 Channels in HEI-OC1 Cells Induced by High Glucose Is Associated with Hearing Impairment

  • Xing, Ying;Ming, Jie;Liu, Tao;Zhang, Nana;Zha, Dingjun;Lin, Ying
    • Yonsei Medical Journal
    • /
    • v.59 no.9
    • /
    • pp.1131-1137
    • /
    • 2018
  • Purpose: Previous reports have shown that hyperglycemia-induced inhibition of transient receptor potential vanilloid sub type 4 (TRPV4), a transient receptor potential ion channel, affects the severity of hearing impairment (HI). In this study, we explored the role of TRPV4 in HI using HEI-OC1 cells exposed to high glucose (HG). Materials and Methods: HEI-OC1 cells were cultured in a HG environment (25 mM D-glucose) for 48 hours, and qRT-PCR and Western blotting were used to analyze the expression of TRPV4 at the mRNA and protein level. TRPV4 agonist (GSK1016790A) or antagonist (HC-067047) in cultured HEI-OC1 cells was used to obtain abnormal TRPV4 expression. Functional TRPV4 activity was assessed in cultured HEI-OC1 cells using the MTT assay and a cell death detection ELISA. Results: TRPV4 agonists exerted protective effects against HG-induced HI, as evidenced by increased MTT levels and inhibition of apoptosis in HEI-OC1 cells. TRPV4 overexpression significantly increased protein levels of phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK), while TRPV4 antagonists had the opposite effect. Our results indicated that TRPV4 is a hyperglycemia-related factor that can inhibit cell proliferation and promote cell apoptosis by activating the MAPK signaling pathway in HEI-OC1 cells. Conclusion: Our results show that the overexpression of TRPV4 can attenuate cell death in HEI-OC1 cells exposed to HG.

Novel Phage Display-Derived H5N1-Specific scFvs with Potential Use in Rapid Avian Flu Diagnosis

  • Wu, Jie;Zeng, Xian-Qiao;Zhang, Hong-Bin;Ni, Han-Zhong;Pei, Lei;Zou, Li-Rong;Liang, Li-Jun;Zhang, Xin;Lin, Jin-Yan;Ke, Chang-Wen
    • Journal of Microbiology and Biotechnology
    • /
    • v.24 no.5
    • /
    • pp.704-713
    • /
    • 2014
  • The highly pathogenic avian influenza A (HPAI) viruses of the H5N1 subtype infect poultry and have also been spreading to humans. Although new antiviral drugs and vaccinations can be effective, rapid detection would be more efficient to control the outbreak of infections. In this study, a phage-display library was applied to select antibody fragments for HPAI strain A/Hubei/1/2010. As a result, three clones were selected and sequenced. A hemagglutinin inhibition assay of the three scFvs revealed that none exhibited hemagglutination inhibition activity towards the H5N1 virus, yet they showed a higher binding affinity for several HPAI H5N1 strains compared with other influenza viruses. An ELISA confirmed that the HA protein was the target of the scFvs, and the results of a protein structure simulation showed that all the selected scFvs bound to the HA2 subunit of the HA protein. In conclusion, the three selected scFVs could be useful for developing a specific detection tool for the surveillance of HPAI epidemic strains.

Experimental studies on the Whitening and Anti-allergic effect of various Herbal medicines that clear heat (수종의 청열약물의 미백 및 항알러지 효과에 대한 실험적 연구)

  • Lee, Myoung-Gyu;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
    • /
    • v.20 no.3
    • /
    • pp.71-81
    • /
    • 2007
  • Objective : In this study, herbal medicine(GJE, Gardenia jasminoides Ellis; HCT, Houttuynia cordata Thunb.; CIL, Chrysanthemum indicum Linne; PMS,Paeonia moutan Sims, P. subfruticosa Makino; APL, Agrimonia pilosa Ledebour) were screened for their inhibitory activities against Tyrosinase and PMA plus A23187-induced $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells to reveal their skin -whitening and anti-allergic effect. Method : To investigate Tyrosinase inhibition we treated Mushroom Tyrosinase(Fluka, 93898) $10{\mu}{\ell}$ and 7.5mM Tyrosine (Sigma, T3754) $20{\mu}{\ell}$ with 80% ethanol medicine extracts. Then we observed 96well micro plate extinction at 490nm. In the next experiment, to investigate Anti-allergic effect we blended cultured Human Mast Cells(HMC-1) with medicine extracts. We treated the blended solution with Phorbol 12-myristate 13-acetate(PMA) and A23187, then observed $TNF-{\alpha}$, IL-6, IL-8 by ELISA (enzyme-linked immunosorbent assay) at 450nm. Results : In inhibiting Tyrosinase the results are as follows. 1. We observed 22% inhibition of Mushroom Tyrosinase at $500{\mu}g/m{\ell}$ concentration of GJE extracts. 2. We also could observe that the decreased Mushroom Tyrosinase activities in HCT, CIL extracts. In inhibiing $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells the results are as follows. 1. Of the extracts examined, HCT, PMS, APL extracts showed over 50% inhibitions of Cytokines at $200{\mu}g/m{\ell}$ concentration. 2. In particular, APL extracts showed the best inhibitory effect on Cytokine productions in a dose-dependent manner. Conclusion : These results suggest that GJE extracts contributes to the anti melanin activities and represent a potential source of whitening agent. Thus these herbal medicines suggest novel drugs on anti-allergic effects.

  • PDF

TIMP-2 Gene Transfer Via Adenovirus Inhibits the Invasion of Lung Cancer Cell (TIMP-2 유전자 재조합 아데노바이러스의 폐암세포 침윤 억제 효과)

  • Oh, Yeon-Mok;Lee, Jae-Ho;Yoo, Chul-Gyu;Chung, Hee-Soon;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
    • /
    • v.49 no.2
    • /
    • pp.189-197
    • /
    • 2000
  • Background : Tissue inhibitor of metalloproteinase is a natural inhibitor that counteracts pro teolytic enzymes essential to the invasion of cancer cell. Whether or not TIMP-2 gene transfer via adenovirus could inhibit the invasion of lung cancer cell iη vitro was evaluated for the future purpose of gene therapy against lung cancer. Methods : Recombinant adenovirus-TIMP-2(Ad-TIMP-2) was generated by homologous recombination after pACCMV-TIMP-2 and pJM17 were cotransfected into 293 cell by standard calcium phosphate coprecipitate method. Calu-6, one of the most invasive lung cancer cells, was transduced with Ad-TIMP-2 or Ad-$\beta$gal. Anchorage-independent growth and invasiveness were assessed by soft agar clonogenicity assay and invasion assay using two-chamber, well divided by matrigel. Results : Ad-TIMP-2 transduced calu-6 cells produced biologically active TIMP-2 more than 50 times more than parental calu-6. TIMP-2 gene transfer did not suppress the in vitro tumorigenicity. However, two chamber well assay revealed that Ad-TIMP-2 transduction reduced the invasiveness of calu-6 efficiently (12% compared with parental cell) even at low 10moi. Conclusion : Even though TIMP-2 gene transfer did not inhibit in vitro tumorigenicity, it did inhibit invasion of lung cancer cell in vitro. The inhibition of invasion by Ad-TIMP-2 may be a useful strategy for the treatment of lung cancer.

  • PDF

Effects of Amomi, Semen Extract on Synthesis of Insulin-like Growth Factor-1 and Anti-wrinkle in Skin (사인추출물의 인슐린 유사 성장인자-3의 합성과 피부 노화 개선에 대한 효과)

  • Choi Gyu Ho;Kim Su Nam;Lee So Hee;Sung Dae Seok;Son Eui Dong;Lee Chang Hoon;Lee Byeong Gon;Jang Ih Seop
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.30 no.3 s.47
    • /
    • pp.389-392
    • /
    • 2004
  • We screened several materials to stimulate IGF-1 promoter activity using luciferase reporter assay and found that Amomi Semen extract (ASE) among them is the most powerful stimulator We also studied about the anti-wrinkle effect of ethanolic extract of Amoni Semen in vitro and in vivo. Semi-quantitative RT-PCR showed that the extract elevated the presence level of IGF-1 mRNA. And $[^3H]$ proline incorporation and semi-quantitative RT-PCR showed that the extract increased the expression of type-I collagen compared with vehicle in vitro and in vivo, respectively. Significant inhibition of MMP-1 expression was determined by ELISA and Western blot. Finally, topical treatment of the extract on hairless mouse's dorsal skin expanded the volume of collagen and dermal thickness. These results suggest that Amomi Semen may be a good candidate for improving extracellular matrix through the increase of collagen expression and inhibition of MMP-1 expression. Moreover, this study enables us to guess that IGF-1 stimulated by the extract may be involved in the mechanism of anti-wrinkle effect of it.

Herbal Extracts as a NF-kappaB Inhibitor (NF-kappaB 프로모터 활성을 억제하는 식물추출물)

  • Park, Deok-Hoon;Lee, Jong-Sung;Jung, Eun-Sun;Hyun, Chang-Gu;Lee, Ji-Young;Hur, Sung-Ran;Koh, Jae-Sook;Lee, Hee-Kyung;Baek, Ji-Hwoon;Yoo, Byung-Sam;Moon, Ji-Young;Kim, Ju-Ho
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.32 no.3 s.58
    • /
    • pp.135-140
    • /
    • 2006
  • Nuclear factor-kappaB (NF-kappaB) is a critical transcription factor for maximal expression of many of the cytokines that are involved in the pathogenesis of inflammatory diseases. In this study, we found that 12 plant extracts among 200 plants, namely, Forsythia koreana, Capsicum annuum L, Mentha arvenis, Duchesnea chrysantha, Morus alba, Saururus Chinenis (Lour) Baill, Pine needle, Zingiber mioga (Thunb.), Roscoe, Houttuynia, Prunus yedoenis, Sasa quelpaertenis, significantly inhibited LPS- induced NF-kappaB activation in a concentration-dependent manner. Additionally, 12 plant extracts were found to have antioxidant activities in DPPH assay Therefore, we have attempted to determine whether 12 herbal extracts could inhibit the expression of cytokines possessing NF-kappaB promoter in their promoter regions. Consistently 12 herbal extracts inhibited LPS-induced production of TNF alpha and interleukin-8 (IL-8). These results show that 12 herbal extracts suppresses the production of pro-inflammatory mediators through the inhibition of the NF-kappaB signaling pathway, we suggest that 12 herbal extracts can be used as a anti-inflammatory and soothing agent.

Quercetin Attenuates the Production of Pro-Inflammatory Cytokines in H292 Human Lung Epithelial Cells Infected with Pseudomonas aeruginosa by Modulating ExoS Production

  • Hye In Ahn;Hyun-Jae Jang;Ok-Kyoung Kwon;Jung-Hee Kim;Jae-Hoon Oh;Seung-Ho Kim;Sei-Ryang Oh;Sang-Bae Han;Kyung-Seop Ahn;Ji-Won Park
    • Journal of Microbiology and Biotechnology
    • /
    • v.33 no.4
    • /
    • pp.430-440
    • /
    • 2023
  • The type three secretion system (T3SS) is a major virulence system of Pseudomonas aeruginosa (P. aeruginosa). The effector protein Exotoxin S (ExoS) produced by P. aeruginosa is secreted into the host cells via the T3SS. For the purpose of an experiment on inhibitors with regard to ExoS secretion, we developed a sandwich-type enzyme-linked immunosorbent assay (ELISA) system. Quercetin was selected because it has a prominent ExoS inhibition effect and also is known to have anti-inflammatory and antioxidant effects on mammalian cells. In this study, we investigated the effects of quercetin on the expression and secretion of ExoS using ELISA and Western blot analysis methods. The results showed that the secretion of ExoS was significantly decreased by 10 and 20 µM of quercetin. Also, popB, popD, pscF, and pcrV which are composed of the T3SS needle, are reduced by quercetin at the mRNA level. We also confirmed the inhibitory effect of quercetin on cytokines (IL-6, IL-1β, and IL-18) in P. aeruginosa-infected H292 cells by real-time polymerase chain reaction (PCR) and ELISA. Collectively, quercetin inhibits the secretion of ExoS by reducing both ExoS production and the expression of the needle protein of T3SS. Furthermore, these results suggest that quercetin has the potential to be used as an anti-toxic treatment for the inflammatory disease caused by P. aeruginosa infection.