STIMULATION OF RAT SKELETAL MUSCLE ANGIOGENESIS BY DIRECT AND CELL-MEDIATED ADMINISTRATION OF RECOMBINANT ANGIOGENIN GENE

Abstract

Objective - to evaluate the effectiveness of revascularization of the rat gastrocnemius muscle following direct and human umbilical cord blood mononuclear cells (MNCs)-mediated delivery of human recombinant angiogenin (Ang) gene to the ischemic area using adenovirus serotype 5 vector (Ad5). Materials and methods. The study was carried out on 30 Wistar rats. Fourteen days after the excision of the femoral artery fragment, the genetic construct was injected into the animals’ ischemic gastrocnemius muscle (AD5-Ang group, n=15). In the other group (mccp+Ad5-Ang, n=15), the transgene was delivered to the muscle with the help of MNCs within the same time limit. In the control group (n=15) 0,9 % NaCl was injected into the muscle of animals under the same conditions. Fourteen and twenty-eight days after the injection, the ratio of capillaries/muscle fibers, the number of muscle fibers and the number of muscle fibers with a central location of nuclei (MCN) were evaluated in the ischemic area. Capillaries were identified by localization of endothelial cells detected by immunohistochemical reaction with antibodies against CD31. Results. On the 14th day after administration of MNCs+Ad5Ang, the ratio of capillaries to the number of muscle fibers in the ischemic area increased by 57 % (p<0,05). On the 28th day in the MNCs+Ad5-Ang group and in the Ad5-Ang group, no significant differences in this indicator were found compared with the control group. The number of muscle fibers on the 14th day in the Ad5-Ang group did not change, and in the MNCs+Ad5-Ang group, it decreased by 58,4 % (p<0,05). By the 28th day, this indicator in the MNCs+Ad5-Ang group decreased by 95,9 % (p<0,05), and in the Ad5-Ang group - by 197,8 % (p<0,05). The number of MCN on the 14th day significantly increased in both experimental groups, in which the genetic constructs were used. Conclusion. The introduction of recombinant ang gene into the area of skeletal muscle ischemia or its delivery to this area with the help of MCNs stimulates angiogenesis and post-ischemic regeneration of muscle fibers.

Keywords

skeletal muscle / ischemia / angiogenesis / angiogenin / mononuclear umbilical cord blood cells / adenoviral vector

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RIGHTS & PERMISSIONS

Samatoshenkov I.V.