TY - JOUR
T1 - Perlecan Heparan Sulfate Proteoglycan Is a Critical Determinant of Angiogenesis in Response to Mouse Hind-Limb Ischemia
AU - Qiang, Beiping
AU - Lim, Sang Yup
AU - Lekas, Michael
AU - Kuliszewski, Michael A.
AU - Wolff, Rafael
AU - Osherov, Azriel B.
AU - Rudenko, Dmitriy
AU - Leong-Poi, Howard
AU - Noyan, Hossein
AU - Husain, Mansoor
AU - Tran, Kiet
AU - Tryggvason, Karl
AU - Hedin, Ulf
AU - Tran-Lundmark, Karin
AU - Strauss, Bradley H.
N1 - Publisher Copyright:
© 2014 Canadian Cardiovascular Society.
PY - 2014/11/1
Y1 - 2014/11/1
N2 - Background: Perlecan is a heparan sulfate proteoglycan (HSPG) constituent of the extracellular matrix with roles in cell growth, differentiation, and angiogenesis. The role of the HS side chains in regulating invivo angiogenesis after hind-limb ischemia is unknown. Methods: Heparan sulfate (HS)-deficient perlecan (Hspg2δ3/δ3) mice (n= 35), containing normal perlecan core protein but deficient in HS side chains, and wild-type (n= 33) littermates underwent surgical induction of hind-limb ischemia. Laser Doppler perfusion imaging (LDPI) and contrast-enhanced ultrasonography (CEU) provided serial assessment of hind-limb perfusion. Harvested muscles underwent immunostaining for endothelial cell density (CD31), real-time reverse transcription polymerase chain reaction RT-PCR for vascular endothelial growth factor (VEGF) mRNA expression and western blot analysis for VEGF and fibroblast growth factor (FGF)2 protein expression at days 2 and28. Results: Serial LDPI showed significantly greater perfusion recovery in ischemic limbs of wild-type compared with Hspg2δ3/δ3 mice. CEU showed that normalized microvascular perfusion was increased in wild-type compared with Hspg2δ3/δ3 mice at day 28 (0.67 ± 0.12 vs 0.26 ± 0.08; P= 0.001). CD31-positive cell counts were significantly higher in wild-type compared with Hspg2δ3/δ3 mice on day 28 (122 ± 30 cells vs 84 ± 34 cells per high-power field [HPF]; P < 0.05). Endogenous VEGF mRNA expression (P < 0.05) and VEGF protein expression (P < 0.002) were significantly decreased in the ischemic limbs of Hspg2δ3/δ3 mice compared with wild-type mice at day 2 and day 28, respectively. FGF2 protein expression showed no significant differences. Conclusions: These results suggest that the HS side chains in perlecan are important mediators of the angiogenic response to ischemia through a mechanism that involves upregulation of VEGF expression.
AB - Background: Perlecan is a heparan sulfate proteoglycan (HSPG) constituent of the extracellular matrix with roles in cell growth, differentiation, and angiogenesis. The role of the HS side chains in regulating invivo angiogenesis after hind-limb ischemia is unknown. Methods: Heparan sulfate (HS)-deficient perlecan (Hspg2δ3/δ3) mice (n= 35), containing normal perlecan core protein but deficient in HS side chains, and wild-type (n= 33) littermates underwent surgical induction of hind-limb ischemia. Laser Doppler perfusion imaging (LDPI) and contrast-enhanced ultrasonography (CEU) provided serial assessment of hind-limb perfusion. Harvested muscles underwent immunostaining for endothelial cell density (CD31), real-time reverse transcription polymerase chain reaction RT-PCR for vascular endothelial growth factor (VEGF) mRNA expression and western blot analysis for VEGF and fibroblast growth factor (FGF)2 protein expression at days 2 and28. Results: Serial LDPI showed significantly greater perfusion recovery in ischemic limbs of wild-type compared with Hspg2δ3/δ3 mice. CEU showed that normalized microvascular perfusion was increased in wild-type compared with Hspg2δ3/δ3 mice at day 28 (0.67 ± 0.12 vs 0.26 ± 0.08; P= 0.001). CD31-positive cell counts were significantly higher in wild-type compared with Hspg2δ3/δ3 mice on day 28 (122 ± 30 cells vs 84 ± 34 cells per high-power field [HPF]; P < 0.05). Endogenous VEGF mRNA expression (P < 0.05) and VEGF protein expression (P < 0.002) were significantly decreased in the ischemic limbs of Hspg2δ3/δ3 mice compared with wild-type mice at day 2 and day 28, respectively. FGF2 protein expression showed no significant differences. Conclusions: These results suggest that the HS side chains in perlecan are important mediators of the angiogenic response to ischemia through a mechanism that involves upregulation of VEGF expression.
UR - http://www.scopus.com/inward/record.url?scp=84908478240&partnerID=8YFLogxK
U2 - 10.1016/j.cjca.2014.06.003
DO - 10.1016/j.cjca.2014.06.003
M3 - Article
C2 - 25249499
AN - SCOPUS:84908478240
SN - 0828-282X
VL - 30
SP - 1444
EP - 1451
JO - Canadian Journal of Cardiology
JF - Canadian Journal of Cardiology
IS - 11
ER -