Intravascular adenovirus-mediated lipoprotein-associated phospholipase A2 gene transfer reduces neointima formation in balloon-denuded rabbit aorta Päivi.

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Intravascular adenovirus-mediated lipoprotein-associated phospholipase A2 gene transfer reduces neointima formation in balloon-denuded rabbit aorta Päivi Turunen, Hanna Puhakka, Juha Rutanen, Mikko O. Hiltunen, Tommi Heikura, Marcin Gruchala, Seppo Ylä-Herttuala Atherosclerosis Volume 179, Issue 1, Pages 27-33 (March 2005) DOI: 10.1016/j.atherosclerosis.2004.10.025 Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 Clinical chemistry analyses. Values (mean±S.E.M.) of plasma Lp-PLA2 activity, total cholesterol, triglycerides, aspartyl aminotransferase (ASAT) and C-reactive protein (CRP) after adenovirus-mediated LacZ or Lp-PLA2 gene transfers. *P<0.05. Atherosclerosis 2005 179, 27-33DOI: (10.1016/j.atherosclerosis.2004.10.025) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 Expression of human Lp-PLA2 mRNA after adenoviral gene transfer in vivo. Lp-PLA2 mRNA was detected by RT-PCR from aortas at two and four weeks time points. The product is 452bp. Line 1: molecular weight marker; line 2: AdLp-PLA2 at two weeks time point; line 3: AdLacZ at two weeks time point; line 4: AdLp-PLA2 at four weeks time point; line 5: AdLacZ at four weeks time point; line 6: positive control (Lp-PLA2 Adeno-X plasmid) and line 7: negative blank control. Atherosclerosis 2005 179, 27-33DOI: (10.1016/j.atherosclerosis.2004.10.025) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 X-gal staining to determine biodistribution of adenoviruses two (A–D) and four weeks (E) after LacZ gene transfer. (A) Aorta; (B) liver; (C) spleen; (D) lung; (E) aorta; (F) Lp-PLA2 transduced aorta; two weeks time point. No staining was detected in heart and kidneys (data not shown). Bars=100μm. Atherosclerosis 2005 179, 27-33DOI: (10.1016/j.atherosclerosis.2004.10.025) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 4 Intima/media ratio (I/M) in the study groups after balloon denudation and gene transfer (mean±S.E.M.). (a) I/M two weeks after the gene transfer. *P<0.005. (b) I/M four weeks after the gene transfer. Atherosclerosis 2005 179, 27-33DOI: (10.1016/j.atherosclerosis.2004.10.025) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 5 Histological characterization of balloon-denuded aortas two weeks after gene transfer. (a–d) LacZ adenovirus transduced aortas. (a) HE-staining; (b) endothelium-specific immunostaining (CD-31); (c) α-actin immunostaining (HHF-35); (d) ApopTag staining. (e–h) Lp-PLA2 adenovirus transduced aortas. (e) HE-staining; (f) endothelium-specific immunostainings (CD-31); (g) α-actin immunostaining (HHF-35); (h) ApopTag staining. Arrowheads denote internal elastic lamina. Magnification×20. Atherosclerosis 2005 179, 27-33DOI: (10.1016/j.atherosclerosis.2004.10.025) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 6 Cell proliferation after balloon denudation and gene transfer. BrdU labelling showed lower proliferation rate in the AdLp-PLA2 group as compared to the AdLacZ group. (a) Two weeks after the gene transfer. (b) Four weeks after the gene transfer. *P<0.05. Atherosclerosis 2005 179, 27-33DOI: (10.1016/j.atherosclerosis.2004.10.025) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions