• 1. Department of Geriatrics, The First Affiliated Hospital, China Medical University, Shenyang 110001, China;;
  • 2. Department of Digestive Internal Medicine, Heilongjiang Province Hospital, Haerbin 150001, China;;
  • 3. Department of General Surgery, The First Affiliated Hospital, China Medical University;;
  • 4. Department of Cardiovascular, The First Affiliated Hospital, China Medical University;;
  • 5. Department of Orthopaedics， Shuangyashan Coal Hospital, Shuangyashan City, Heilongjiang 155100, China;
Bone mesenchymal stem cells; 　　Enhanced green fluorescent protein; 　　Akt gene; 　　Hindlimb ischemic　　Angiogenesis
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Objective　To investigate the effect of bone mesenchymal stem cells (MSCs) transfected with pEGFP-C1/Akt after intramuscular injection on angiogenesis in ischemic hindlimb of mice. Methods　A total of 30 Wistar mice used in the present experiments were divided into gene therapy group (injected MSCs transfected with pEGFP-C1/Akt), non-gene therapy group （injected MSCs） and control group (injected PBS). Mice skin temperature in left hindlimbs was measured by a infrared ray temperature meter before and right after ischemia and 1-7 days after transfection, respectively. Digital subtraction angiography was carried out to observe angiogenesis in hindlimb of mice after 28 days. The capillary density was determined with immunohistochemical analysis in the resected specimen. The expressions of Akt mRNA and protein, VEGF mRNA and protein were measured via RT-PCR and Western blot. Results　Skin temperature increased significantly on 3 d after transfection in gene therapy group. On 28 d after transfection， angiography showed that the number of collateral vessels markedly increased in gene therapy　group. Green fluorescence cells were observed in ischemic tissue from the adductor and semimembranous muscles of gene therapy group under fluorescent microscope. The result of immunohistochemical analysis of capillary density showed that the capillary density was significantly higher in gene therapy group 〔(7.1±0.3)/high power〕 than that in non-gene therapy group 〔(4.2±0.4)/high power〕 and control group 〔(1.3±0.2)/high power〕, with significantly statistical differences among three groups (P＜0.01). Akt mRNA (2.44±0.14) and protein (1.1±0.13), VEGF mRNA (1.1±0.11) and protein (0.97±0.13) of gene therapy group markedly increased compared with Akt mRNA (1.58±0.13) and protein (0.78±0.12), VEGF mRNA (0.78±0.14) and protein (0.67±0.11) of non-gene therapy group as well as Akt mRNA (0.64±0.11) and protein (0.36±0.12), VEGF mRNA (0.56±0.11) and protein (0.33±0.13) of control group (P＜0.01). Conclusion　The effect of MSCs transfected with pEGFP-C1/Akt on angiogenesis is significantly better than only MSCs therapy, which may offer a new way for the ischemic diseases.

Citation： HUO Xin,FENG Jinwei,LIU Fangfeng,KONG Hongliang,YAN Yongji,ZHANG Qiang. Effect of Bone Mesenchymal Stem Cells Transfected with pEGFP-C1/Akt on Angiogenesis in Ischemic Hindlimb of Mice. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2008, 15(6): 402-407. doi: Copy