Preventive Effection Of Gene Therapy With ENOS Gene On The Formation Of Thrombi Of Vascular Anastomotic Site

Objective To identify the effects of in vivo local expression of recombine eNOS gene on the thrombosis formation in vascular anastomotic site.Methods 30 SD rats were divided into treatment group(n=15)and control group (n=15). End to end anastomoses were performed in rat femoral arteries. Lipofection 2000 and recombinant pcDNA3.0-eNOS(treatment group)or Lipofection 2000 and pcDNA3.0(control group)were injected into anastomotic vascular lumen for 30 minutes. 5 rats of each group were executed 5 days after the operation to obtain 1.5 cm length vascular tissue. The vascular tissue were divided to 3 portiones equally. The expression of eNOS gene in vascular anastomotic site was detected by RT-PCR and Western blotting assay. The NO activity was measured by chemiluminescent assay. Other rats were executed 2 weeks after the operation to obtain 0.5 cm length vascular tissue for pathological study.RESULTS When the isolated RNA was performed with RT-PCR, 591bp band appeared in the treatment group, but band could not be found in the control group. The eNOS and NO in treatment group were significantly higher than those in control group. Thrombosis rate fewer in the treatment group than that in the control group (P<0.01).Conclusion When pcDNA3.0-eNOS were transferred into anastomotic vascular wall, the foreign gene can produce eNOS at anastomotic sites and effectively prevent the formation of thrombosis.