Chemical analysis of atherosclerotic plaque cholesterol combined with histology of the same tissue
| Autor: | Robert M. Christie, J. Michael Kalbfleisch, Ming-Shang Kuo, Donetta Gifford-Moore, Pamela Rutherford, Mark David Rekhter, Kwan Hui, Kenneth E. Gould, Xiao-di Huang |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2008 |
| Předmět: |
Genetically modified mouse
Agonist Pathology medicine.medical_specialty medicine.drug_class QD415-436 transgenic mice Biochemistry Lesion Mice chemistry.chemical_compound Endocrinology Antigen Tandem Mass Spectrometry medicine Animals liquid chromatography Derivatization Chromatography High Pressure Liquid mass spectrometry Mice Knockout Chromatography Chemistry Cholesterol Histology Cell Biology Reference Standards Immunohistochemistry quantification animal models medicine.symptom atherosclerosis |
| Zdroj: | Journal of Lipid Research, Vol 49, Iss 6, Pp 1353-1363 (2008) |
| ISSN: | 0022-2275 |
| Popis: | Sensitive method for chemical analysis of free cholesterol (FC) and cholesterol esters (CE) was developed. Mouse arteries were dissected and placed in chloroform-methanol without tissue grinding. Extracts underwent hydrolysis of cholesteryl esters and derivatization of cholesterol followed by liquid chromatography/mass spectrometry (LC/MS/MS) analysis. We demonstrated that FC and CE could be quantitatively extracted without tissue grinding and that lipid extraction simultaneously worked for tissue fixation. Delipidated tissues can be embedded in paraffin, sectioned, and stained. Microscopic images obtained from delipidated arteries have not revealed any structural alterations. Delipidation was associated with excellent antigen preservation compatible with traditional immunohistochemical procedures. In ApoE(-/-) mice, LC/MS/MS revealed early antiatherosclerotic effects of dual PPARalpha,gamma agonist LY465606 in brachiocephalic arteries of mice treated for 4 weeks and in ligated carotid arteries of animals treated for 2 weeks. Reduction in CE and FC accumulation in atherosclerotic lesions was associated with the reduction of lesion size. Thus, a combination of LC/MS/MS measurements of CE and FC followed by histology and immunohistochemistry of the same tissue provides novel methodology for sensitive and comprehensive analysis of experimental atherosclerotic lesions. |
| Databáze: | OpenAIRE |
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