Quantifying Myocardial Oxidative Stress and Microvasculature

Myocardial in situ production of superoxide anion was detected by dihydroethidium (DHE) staining (20μM/l; Sigma) [7 (link)], and myocardial capillary density was determined by CD31 staining (Bio-Rad, Oxford, UK). In addition, myocyte cross-sectional area and myocardial fibrosis were calculated in wheat germ agglutinin (WGA)- and trichrome-stained slides, respectively. All staining images were semi-automatically quantified in 15–20 fields and the results from all fields averaged [29 (link)]. Myocardial microvascular structure was assessed by micro-CT, as previously described [24 (link)]. The LV wall was perfused under physiological pressure with an intravascular contrast agent (MV-122; Flow Tech, Carver, MA) through a branch of the left circumflex coronary artery. A transmural portion (2 cm³) of the LV was scanned, and images were analyzed with Analyze™. Spatial density of microvessels (20-500μm) in the subepicardium and subendocardium and vessel tortuosity (an index of vessel immaturity) were calculated, as previously described [24 (link)].

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Farahani R.A., Yu S., Ferguson C.M., Zhu X.Y., Tang H., Jordan K.L., Saadiq I.M., Herrmann S.M., Chade A.R., Lerman A., Lerman L.O, & Eirin A. (2021). Renal Revascularization Attenuates Myocardial Mitochondrial Damage and Improves Diastolic Function in Pigs with Metabolic Syndrome and Renovascular Hypertension. Journal of cardiovascular translational research, 15(1), 15-26.

Publication 2021

dihydroethidium (DHE) MV-122

Capillary Contrast agent Coronary artery Dihydroethidium Fibrosis Micro ct Myocardial Myocyte Physiological Pressure Superoxide anion Vessel Wheat germ agglutinin

Corresponding Organization :

Other organizations : Mayo Clinic, University of Mississippi Medical Center, Jackson Memorial Hospital, Mayo Clinic in Arizona

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Variable analysis

independent variables

Myocardial in situ production of superoxide anion

dependent variables

Myocardial capillary density
Myocyte cross-sectional area
Myocardial fibrosis
Spatial density of microvessels (20-500μm) in the subepicardium and subendocardium
Vessel tortuosity (an index of vessel immaturity)

control variables

Physiological pressure for perfusion of the LV wall with an intravascular contrast agent

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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