[Gene transfers of kallikrein, bradykinin receptor B2 and a mutated B2 receptor stimulate neoangiogenesis in peripheral ischemia].
In this work, we evaluated the angiogenic effect of the gene transfer of human tissue kallikrein (TK), bradykinin B2 receptor (B2R) and a mutated form (RB2m) in a rabbit peripheral model of ischaemia. We studied the effects of the transfection of each of these factors and the effects of their co-transfection. In New Zealand anesthetised rabbits we first induced an ischaemia of the left posterior leg by ligation-excision of the superficial femoral artery and its collaterals. Seven days later, we performed i.m. injections in the ischemic tight with transfection solutions containing either the control (pcDNA3 empty backbone) or the pcDNA3-TK, the pcDNA3-TK and the pcDNA3-B2R, the pcDNA3-TK and the pcDNA3-B2Rm. Twenty eight days later, the therapeutic effect was evaluated using ultrasonographic debitmetry of the common iliac artery, perfusion index (PI) = ischemic leg blood flow /non ischemic leg blood flow (%) and capillaries measurements i.e. capillary density: number of vessels/mm2 and the ratio of vessels/muscular fiber, in the adductors and gastrocnemian muscles. The PI was increased in each treated group vs control (32.61 +/- 5.2%), pcDNA3-TK: 59.72 +/- 2.33%; p = 0.001; pcDNA3-RB2: 55.25 +/- 2.29%; p = 0.008; pcDNA3-TK + pcDNA3-RB2: 84.77 +/- 3.15%; p < 0.001; pcDNA3-TK + pcDNA3-RB2m: 103.25 +/- 4.9%; p < 0.001. The capillary density and the vessel/muscular fiber ratio increased in a parallel with the hemodynamic in the ischemic adductors (pcDNA3-TK + pcDNA3-B2Rm > pcDNA3-TK + pcDNA3-RB2 > pcDNA3-TK = pcDNA3-B2R; p < 0.001). There was no angiogenic effect measurable neither in the non ischemic adductors (right) nor in the gastrocnemian muscles. In rabbit peripheral ischaemia, the cotransfection of TK and B2R increases the arterial flow in the treated leg and potentiates the neoangiogenesis. Cotransfection of the B2Rm cDNA enhanced the synergic effect of this therapeutic strategy.
Griffon
C
C
Laboratoire de recherche sur l'angiogenèse, Université Louis Pasteur, Strasbourg.
Boehm
N
N
Allegrini
J
J
Miternique
A
A
Alhenc-Gelas
F
F
Hudlett
P
P
Stephan
D
D
D000818:Animals; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D006614:Hindlimb / Q000098:blood supply; D007511:Ischemia / Q000235:genetics / Q000628:therapy* / Q000662:veterinary*; D007610:Kallikreins / Q000235:genetics* / Q000502:physiology*; D009154:Mutation; D018919:Neovascularization, Physiologic / Q000235:genetics*; D016491:Peripheral Vascular Diseases / Q000235:genetics / Q000628:therapy / Q000662:veterinary; D011817:Rabbits; D043782:Receptor, Bradykinin B2 / Q000235:genetics* / Q000502:physiology*; D012039:Regional Blood Flow; D014162:Transfection
D043782:Receptor, Bradykinin B2; D007610:Kallikreins