Micro ATR-FTIR spectroscopic imaging of atherosclerosis: an investigation of the contribution of inducible nitric oxide synthase to lesion composition in ApoE-null mice.
Inducible nitric oxide synthase (iNOS) has previously been shown to contribute to atherosclerotic lesion formation and protein nitration. Micro attenuated total reflection (ATR)-Fourier transform infrared (FTIR) spectroscopic imaging was applied ex vivo to analyse lesions in atherosclerotic (ApoE-/-) mice. Histologies of cardiovascular tissue of ApoE-/- mice that contain the gene for iNOS and ApoE-/- mice without iNOS (ApoE-/-iNOS-/- mice) were examined. Spectroscopic imaging of the aortic root revealed that iNOS did not affect the composition of the tunica media; furthermore, irrespective of iNOS presence, lipid esters were found to form the atherosclerotic plaque. ApoE-/- mouse aortic root lesions exhibited a more bulky atheroma that extended into the medial layer; signals characteristic of triglycerides and free fatty acids were apparent here. In ApoE-/-iNOS-/- mouse specimens, lesions composed of free cholesterol were revealed. ATR-FTIR spectra of the intimal plaque from the two mouse strains showed higher lipid concentrations in ApoE-/- mice, indicating that iNOS contributes to lesion formation. The reduction of lesion prevalence in ApoE-/-iNOS-/- mice compared with ApoE-/- mice is consistent with previous data. Moreover, the analysis of the plaque region revealed a change in the spectral position of the amide I band, which may be indicative of protein nitration in the ApoE-/- mouse, correlating with a more ordered (beta-sheet) structure, while a less ordered structure was apparent for the ApoE-/-iNOS-/- mouse, in which protein nitration is attenuated. These results indicate that micro ATR-FTIR spectroscopic imaging with high spatial resolution is a valuable tool for investigating differences in the structure and chemical composition of atherosclerotic lesions of ApoE-/- and ApoE-/-iNOS-/- mice fed a high-fat Western diet and can therefore be applied successfully to the study of mouse models of atherosclerosis.
Palombo
Francesca
F
Department of Chemical Engineering, Imperial College London, London, UK SW7 2AZ.
Shen
Hao
H
Benguigui
Lea Esther S
LE
Kazarian
Sergei G
SG
Upmacis
Rita K
RK
D000818:Animals; D001011:Aorta / Q000378:metabolism; D001057:Apolipoproteins E / Q000172:deficiency* / Q000235:genetics*; D050197:Atherosclerosis / Q000175:diagnosis / Q000378:metabolism* / Q000473:pathology* / Q000503:physiopathology; D003952:Diagnostic Imaging; D004041:Dietary Fats; D017353:Gene Deletion; D006801:Humans; D008297:Male; D051379:Mice; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared; D013997:Time Factors; D017539:Tunica Intima / Q000378:metabolism
D001057:Apolipoproteins E; D004041:Dietary Fats; D052247:Nitric Oxide Synthase Type II