Journal: American Journal of Physiology - Heart and Circulatory Physiology
Article Title: Endothelial cells overexpressing IL-8 receptor reduce cardiac remodeling and dysfunction following myocardial infarction
doi: 10.1152/ajpheart.00571.2012
Figure Lengend Snippet: Effects of IL8RA/RB-EC, Null-EC, or vehicle transfusion on neutrophil and monocyte/macrophage infiltration into myocardial infarction (MI) injured heart 24 h after transfusion of ECs. Sham-operated (no MI) control rats were age-matched male Sprague-Dawley rats without MI or EC transfusion. Vehicle rats were MI rats that did not receive EC transfusion. IL8RA/RB-EC or Null-EC rats were MI rats that received intravenous transfusions of Null-ECs or ECs overexpressing IL8RA and IL8RB (IL8RA/RB-ECs, 0.5 × 106 cells/injection in 500 μl saline) at 1, 3, and 5 h post-left anterior descending (LAD) coronary artery ligation. Null-ECs were ECs transduced with the empty adenoviral vector. Neutrophils and monocytes/macrophages were immunostained with MPO and ED1 primary antibodies, respectively, on 5 μm cross sections of left ventricle (LV) 2 mm below the LAD coronary artery ligation. A and B: representative micrographs showing the MPO+ and ED1+ cells in the area at risk (within 2 mm adjacent to the infarct region) of injured hearts, respectively. C and D: bar graphs showing neutrophil and monocyte/macrophage densities in the area at risk of injured hearts of rats transfused with vehicle, Null-ECs, or IL8RA/RB-ECs. Results are means ± SE; (n) = number of rats. One-way ANOVA was used to analyze the data. *P < 0.05 vs. respective sham groups; #P < 0.05 vs. respective MI + vehicle and MI + Null-EC groups.
Article Snippet: Rat aortic ECs (purchased from VEC Technologies, Cat No. RAEC/T-75) that overexpress IL8RA-GFP, IL8RB-GFP, or Null-GFP (empty adenoviral vector control) were generated using adenoviral vectors that contain human IL8RA and/or IL8RB cDNAs and the GFP gene using the AdEasy Adenoviral Vector System (Stratagene) as reported previously ( 35 ).
Techniques: Injection, Ligation, Transduction, Plasmid Preparation
![Transfusion of IL8RA/RB-ECs increased capillary density in the area at risk post-MI. A: representative micrographs (400×) of cross sections (frozen sections) of the area at risk of MI + Vehicle, MI + Null-EC, and MI + IL8RA/RB-EC rat hearts double immunostained with CD31 (red color, capillary EC marker) and GFP (green color, IL8RA/RB-EC marker) 24 h post-MI. ECs stained with both CD31 and GFP appear yellow in color. Nuclei stained with 4,6-diamidino-2-phenylindole (DAPI) were blue in color. B: representative micrographs (400×) of cross sections (formalin-fixed, paraffin-embedded) of an MI-injured LV immunostained with CD31 (brown color) 2 wk post-MI. C: bar graphs showing capillary densities [ratios of CD31+ area vs. total tissue area per microscopic field (400×)] in myocardium of sham-operated (no MI surgery), MI + Vehicle, MI + Null-EC, and MI + IL8RA/RB-EC rats 2 wk post-MI. Remote area was uninjured area in septum; area at risk was injured area in posterior wall (border zone surrounding the infarct region, using the collagen-stained slice as a reference). Twenty fields of area at risk and 10 fields of remote area were sampled from each rat heart. In the sham-operated group the hatched bar represents capillary density in the uninjured posterior wall. Results are means ± SE; (n) = number of hearts. *P < 0.05 vs. respective remote area; #P < 0.05 vs. respective MI + Vehicle and MI + Null-EC groups.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_1247/pmc03891247/pmc03891247__zh40161308330006.jpg)
