| Optimization of cell-based assays to quantify the anti-inflammatory/allergic potential of test substances in 96-well format. | |
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MedLine Citation:
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PMID: 21069571 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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OBJECTIVE: There is an insistent need for robust, reliable, and optimized assays for screening novel drugs targeting the inflammatory/allergic markers. The present study describes about the optimization of eight cell-based assays utilizing mammalian cell lines in 96-well format for quantifying anti-inflammatory/allergic drug candidates. MATERIALS AND METHODS: We estimated the inhibitory response of reference compounds: 1400W dihydrochloride on LPS-induced NO release, celecoxib on LPS-induced PGE(2) production and dexamethasone on LPS-induced pro-inflammatory cytokines IL-1 beta, IL-6, and TNF-alpha production by J774A.1 murine macrophages. Response of acetylsalicylic acid and celecoxib was studied on A23187-induced TXB(2) production; captopril on A23187-stimulated LTB(4) production by HL-60 cells. Effect of ketotifen fumarate was evaluated on A23187-elicited histamine release by RBL-2H3 cells. Each experiment was repeated twice to assess the reproducibility and suitability of the assays by determining appropriate statistical tools viz. %CV, S/B and Z' factor. RESULTS: 1400W dihydrochloride was capable of inhibiting LPS-induced NO levels (IC(50) = 10.7 μM). Dexamethasone attenuated LPS-induced IL-1 beta (IC(50) = 70 nM), IL-6 (IC(50) = 58 nM) and TNF-alpha (IC(50) = 44 nM) release, whereas celecoxib, a specific COX-2 inhibitor showed marked reduction in LPS-induced PGE(2) (IC(50) = 23 nM) production. Captopril (IC(50) = 48 μM) and ketotifen fumarate (IC(50) = 36.4 μM) demonstrated potent inhibitory effect against A23187-stimulated LTB(4) and histamine levels, respectively. Both acetylsalicylic acid (IC(50) = 5.5 μM) and celecoxib (IC(50) = 7.9 nM) exhibited concentration-dependent decrease in TXB(2) production. Results for all the cell assays from two experiments showed a Z' factor varying from 0.30 to 0.99; the S/B ratio ranged from 2.39 to 24.92; %CV ranged between 1.52 and 20.14. CONCLUSION: The results proclaim that these cell-based assays can act as ideal tools for screening new anti-inflammatory/anti-allergic compounds. |
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Authors:
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C V Chandrasekaran; R Edwin Jothie; Preeti Kapoor; Anumita Gupta; Amit Agarwal |
Publication Detail:
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Type: Journal Article Date: 2010-10-31 |
Journal Detail:
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Title: Inflammopharmacology Volume: 19 ISSN: 1568-5608 ISO Abbreviation: Inflammopharmacology Publication Date: 2011 Jun |
Date Detail:
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Created Date: 2011-05-10 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9112626 Medline TA: Inflammopharmacology Country: Switzerland |
Other Details:
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Languages: eng Pagination: 169-81 Citation Subset: IM |
Affiliation:
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Department of Cellular Assay, R&D Centre, Natural Remedies Pvt. Ltd, Plot No. 5B, Veerasandra Industrial Area, 19th K. M. Stone, Hosur Road, Bangalore, 560100, India, cvc@naturalremedy.com. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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