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Rescue of hypercholesterolemia-related impairment of angiogenesis by oral folate supplementation

Ken-ichiro Sasaki, MD^*, Junli Duan, MD^*, Toyoaki Murohara, MD^*,,*, Hisao Ikeda, MD^*, Satoshi Shintani, MD^*, Toshifumi Shimada, MD^*, Takako Akita, MD^*, Kimiyasu Egami, MD^* and Tsutomu Imaizumi, MD, FACC^*

^* Cardiovascular Research Institute, Kurume University School of Medicine, Kurume, Japan
Department of Cardiology, Nagoya University Graduate School of Medicine, Nagoya, Japan

View larger version (51K): [in a new window]   Figure 1 (A) Representative laser Doppler blood flow (LDBF) images at days 7, 14, and 28 are shown. (B) Before and after surgery, the hindlimb LDBF ratio did not differ among the three groups. The control group revealed progressive recovery of the ischemic/normal LDBF ratio within 28 days after induction of limb ischemia. However, the LDBF ratio was lower in the hypercholesterolemia (HC) group than in the controls. In the HC + Folate group, the LDBF ratios were significantly improved compared with the HC group; n = 6 in each group. *p < 0.05; **p < 0.01 vs. control; p < 0.05; p < 0.01 vs. HC.

View larger version (46K): [in a new window]   Figure 2 (A) Representative postmortem angiograms taken at day 14 are shown. There are numerous collateral vessels in the medial thigh area in the control group. However, the hypercholesterolemia (HC) group showed reduced collateral vessels. Folate supplementation (HC + Folate group) increased collateral vessels. (B) The angiographic score was significantly lower in the HC than in the control group, which was rescued in the HC + Folate group. n = 6 in each group; ***p < 0.001.

View larger version (33K): [in a new window]   Figure 3 (A) The capillary density was significantly lower in the hypercholesterolemia (HC) group than in the control group in the ischemic hindlimb, which was restored in the HC + Folate group. In the nonischemic hindlimb, the capillary densities did not differ among the three groups. (B) Similarly, the capillary/muscle fiber ratio was significantly lower in the HC group than in the control group in the ischemic hindlimb, which was restored in the HC + Folate group. In the nonischemic hindlimb, the capillary/muscle fiber ratio did not differ among the three groups. n = 6 in each group; ***p < 0.001.

View larger version (19K): [in a new window]   Figure 4 The effects of nitric oxide inhibition on angiogenesis. (A) Before and after surgery, the ischemic/normal laser Doppler blood flow (LDBF) ratio did not differ among the three groups. The reduced LDBF ratio in the hypercholesterolemia (HC) group was restored by folate administration (HC + Folate group). Treatment with N-nitro-L-arginine methyl ester (L-NAME), the nitric oxide synthase inhibitor, markedly suppressed the recovery of LDBF ratio (HC + Folate + L-NAME group). n = 6 in each group; **p < 0.01 vs. HC + Folate; p < 0.05; p < 0.01 vs. HC. (B) The angiographic score was greater in the HC + Folate group than in the HC group. The score was significantly suppressed by L-NAME treatment (HC + Folate + L-NAME group). n = 6 in each group **p < 0.01; ***p < 0.001.