| Assay of mitochondrial functions by resazurin in vitro. | |
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MedLine Citation:
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PMID: 15000895 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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AIM: To study the mechanism of resazurin as indicator of mitochondrial function and to develop a rapid and sensitive assay for measuring metabolic activity of isolated mitochondria from rat liver in vitro. METHODS: The screening was carried out on 96-well microtitre plates by monitoring fluorescence intensity of resazurin reduced by mitochondria. Experimental conditions were optimized and influences of several inhibitors on mitochondrial function were observed. RESULTS: Fluorescence intensity increased in a linear manner when the mitochondrial protein concentration from 5 to 50 microg protein per well was incubated with resazurin (5 micromol/L) during 230 min period at 37 degrees C. Edetic acid could promote the reduction of resazurin in mitochondria. The fluorescence intensity decreased greatly after pretreatment with NaN3, antimycin A, carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), and oligomycin compared with the control. However, the typical complex I inhibitor, rotenone enhanced the fluorescence intensity without mitochondria. CONCLUSION: Using resazurin to determine mitochondrial function is sensitive, inexpensive and could be easily automated for high throughput screening. |
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Authors:
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Hai-xia Zhang; Guan-hua Du; Jun-tian Zhang |
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Publication Detail:
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Type: In Vitro; Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Acta pharmacologica Sinica Volume: 25 ISSN: 1671-4083 ISO Abbreviation: Acta Pharmacol. Sin. Publication Date: 2004 Mar |
Date Detail:
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Created Date: 2004-03-05 Completed Date: 2004-08-06 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 100956087 Medline TA: Acta Pharmacol Sin Country: China |
Other Details:
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Languages: eng Pagination: 385-9 Citation Subset: IM |
Affiliation:
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Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100050, China. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Antimycin A / pharmacology Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / pharmacology Edetic Acid / pharmacology Fluorescence Male Mitochondria, Liver / drug effects, metabolism* Mitochondrial Proteins / metabolism Nitrates / pharmacology Oligomycins / pharmacology Oxazines / metabolism* Rats Rats, Sprague-Dawley Rotenone / pharmacology Xanthenes* |
| Chemical | |
Reg. No./Substance:
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0/Mitochondrial Proteins; 0/Nitrates; 0/Oligomycins; 0/Oxazines; 0/Xanthenes; 370-86-5/Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; 550-82-3/resazurin; 60-00-4/Edetic Acid; 642-15-9/Antimycin A; 7631-99-4/sodium nitrate; 83-79-4/Rotenone |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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