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Method for detecting a biomarker of oxidative stress in a biological sample

A biomarker and biological sample technology, applied in the field of detection of oxidative stress in biological samples, can solve problems that have not been described

Inactive Publication Date: 2009-03-04
INST DE CARDIOLOGIE DE MONTREAL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the possibility that circulating HNE-protein adducts may reflect heightened systemic or tissue-specific oxidative stress, and methods for the quantification of these adducts in whole blood, plasma, or other blood derivative samples, has not been previously examined. , has not been described before

Method used

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  • Method for detecting a biomarker of oxidative stress in a biological sample
  • Method for detecting a biomarker of oxidative stress in a biological sample
  • Method for detecting a biomarker of oxidative stress in a biological sample

Examples

Experimental program
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Embodiment 1

[0051] Conduct an assay to quantify NaB 2 h 4 HNE and its inactive metabolite 1,4-dihydroxynonene (DHN) bound to thiol protein adducts after Raney nickel treatment. The levels of these adducts were measured in blood, plasma collected from 7, 15, 22 and 30 week old SHR and control Wistar rats. Disease (SHR) and age (p<0.0001 for both) significantly increased the levels of protein-bound HNE quantified at nanomolar levels with relatively high accuracy in blood but not plasma. Compared with Wistar rats, 22- and 30-week-old SHRs showed higher blood levels of HNE-protein adducts. Protein-bound DHN levels detected in blood and plasma were not affected by disease or age. Taken together, the results of this study in an animal model of cardiomyopathy demonstrate that changes in blood HNE-protein thioether adducts with disease progression and aging can be assessed with high accuracy by the described GCMS method. It is expected that this method will be used to assess the occurrence an...

Embodiment 2

[0053] Another experiment was performed to evaluate the role of 4-hydroxynonenal (HNE) in oxidative stress-related diseases. Further, based on the discovery of high circulating HNE-protein thioether adducts (HNE-P) in essential hypertensive rats (SHR), the purpose of this study was to find out the correlation between HNE-P and cardiac function properties and to examine the effects of antioxidant treatment.

[0054] Eighteen-week-old SHRs (9 rats / group) were administered daily (i.p.) the lipid peroxidation inhibitor probucol (10 mg / kg / day) or vehicle (corn oil) for 4 weeks. Cardiac function was assessed by echocardiography and HNE-P by GCMS.

[0055]Changes (p<0.05) in left ventricular diastolic (increased isovolumic relaxation time) and compliance (increased E-wave deceleration rate, EDR) indices reflected worsening diastolic dysfunction in SHRs receiving vehicle. High circulating HNE-P was associated with diastolic dysfunction (EDR: R2=0.518; p<0.001) and heart rate (R2=0.2...

Embodiment 3

[0057] In the next example, the method for the determination of protein-bound HNE in myocardial tissue using quantitative GCMS [18] was modified to enable accurate and reproducible continuous assessment of low levels of these adducts in blood samples. Specifically, this method quantifies HNE and its inactive metabolite DHN bound to thiol proteins. However, in alternative embodiments of the invention, oxidative stress may be quantified with any other suitable substance, such as any other metabolite of an aldehyde produced by fatty acid peroxidation. HNE-protein thioether adducts and DHN-protein thioether adducts are considered representative of the class of aldehyde-protein adducts and aldehyde metabolite-protein adducts.

[0058] In general, the method involves: (i) by reduction to its deuterated alcohol [ 2 H]DHN to stabilize HNE, (ii) treatment with Raney nickel to cleave the thioether bond, releasing protein-bound DHN and [ 2 H]DHN(HNE), and (iii) using deuterated interna...

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Abstract

The present invention relates to methods for detecting a oxidative stress in a biological sample, methods of determining a cumulative record of oxidative injury, and methods of diagnosing diseases of aging, such as cardiovascular diseases, based on the presence or absence of a biomarker or a component thereof. The present invention also relates to a kit for detecting oxidative stress in a biological sample comprising a stabilizing reactant and an antibody.

Description

technical field [0001] The present invention relates to methods for detecting oxidative stress in biological samples, methods for determining the cumulative profile of oxidative damage, and methods for diagnosing diseases of aging, such as cardiovascular disease, based on the presence or absence of biomarkers or components thereof. The invention also relates to a kit for detecting oxidative stress in a biological sample comprising a stabilizing reagent and an antibody. Background technique [0002] Over the past 30 years, a wealth of experimental evidence supporting the association of oxidative stress with aging and the onset of cardiovascular disease (CVD) has accumulated [1-3]. The notoriously disappointing randomized clinical trials of "natural antioxidants" (HOPE, HPS, GISSI-prevention) have led some to question the relevance of the oxidative stress hypothesis [4-6]. However, most of these studies did not evaluate the effect of antioxidant interventions on oxidative str...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53A61K31/00A61K31/10A61P9/00G01N33/483G01N33/50
Inventor 克里斯廷·迪斯－罗西尔斯卡罗琳·阿斯兰伯特兰·布查德吉恩－克劳德·塔迪芙布兰丁·孔德
Owner INST DE CARDIOLOGIE DE MONTREAL
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