Synergistic Role Of Astragalus Mongholicus And/or Mailuoning Injection With Modified Collagen In Enhancing Angiogensis

[Background] Tissue engineering synthesized the artificial tissues and organs to find a method for rapid vascularization in stent. Angiogenesis and plays an important role in tissue repairing and promoting cell growth. Studies have shown that astragalus mongholicus, Mailuoning may have some role in promoting angiogenesis, and the mechanism may be promoting vascular endothelial cell proliferation and DNA synthesis, inhibition of apoptosis, raising a variety of angiogenic growth factors such as vascular endothelial growth factor, platelet derived growth factor and basic fibroblast growth factor and so the expression of cell hemoglobin and increased hydroxyproline ammonia acid content. When we put the Astragalus mongholicus and Angelica with ratio of 1:5 in the CAM, the results show that angiogenesis and blood vessel counts, the ratio was significantly better than the other group and saline control group, with better promotion of angiogenesis in CAM. Although these experiments show different aspects of traditional Chinese medicine with the optimization process of the body to promote the role of vascular endothelial growth factor, but current research were all confined to oral and intravenous administration, the previous research has confirmed Astragalus mongholicus microemulsion modified-collagen in promoting the role of CAM angiogenesis with a similar effect of VEGF.[Objective] This study loaded the Astragalus mongholicus and mailuoning in modified collagen, to investigate the efficacy of CAM angiogenesis, and prove whether there was synergy of Astragali and Mailuoning, and to explore the angiogenic effects and possible mechanisms of modified collagen matrices loading with Astragalus Mongholicus-Maluoning injection with cell experiments.[Methods] The Astragalus mongholicus injection, Mailuoning injection and their mixer were added heparin sodium-modified collagen (type I collagen). Five groups were set in the experiment, including blank group without any interfere, control group with only modified collagen implanted into chorioallantoic membrane, Astragalus mongholicus group with 1 mL of Astragalus mongholicus injection loading in collagen, Mailuoning group with 1 mL of Mailuoning injection loading in collagen, Astragalus mongholicus plus Mailuoning group with 0.5 mL Astragalus mongholicus and 0.5 mL Mailuoning injection loading in collagen. After 30 mintutes they were implanted for 7 days, the samples were explanted from the chorioallantoic membrane. Record the number of embryo survival, chorioallantoic membrane vascular growth shape, chorioallantoic membrane wrapped specimens into the extent, the number of micro-vessels in chorioallantoic membrane was counted at 50 times the magnification of the microscope with not repeat the three horizons capillary method using grid, the hemoglobin was determined by photometric, while taking the wound center granulation tissue, CD31 immunohistochemistry were done by the pathologist to cout the rhVEGF165 positive cells in three selected vision randomly. Two groups of collagen matrices loading with or without Astragalus Mongholicus-Mailuoning injection were set in the experiments, each of which was cultured with thoracic aorta endothelial cells for 24 hours. Afterwards, the effects of cell proliferation, migration and tube formation were detected followed by protein expression of VEGF with western blot assay.[Results] For the appearance of macro-samples, the rate of wheel spoke-like vessels and enclosing area of samples by chorioallantoic membrane in each experimental group were better than blank group and control group. The number of micro-vessels in the chorioallantoic membrane, together with hemoglobin content and the number of rhVEGF165 positive cells in the collagen matrices were significantly higher in each experimental group than those in the control group (P< 0.01). Among the experimental groups, the index of Astragalus mongholicus plus Mailuoning group was significantly higher than the results of Astragalus mongholicus alone and Mailuoning alone groups (P< 0.05). The group of collagen matrices loading with Astragalus Mongholicus-Mailuoning injection can positively increase rat thoracic aorta endothelial cell proliferation to 1.9022 times, migration to 2.1143 times and tube formation to 2.46 times as compared to the results of group of collagen without loading Astragalus Mongholicus-Mailuoning injection. The protein expression of VEGF improved 0.638 time in the group of collagen matrices loading with Astragalus Mongholicus-Mailuoning injection as compared to that without loading.[Conclusion] Loading Astragalus mongholicus and/or Mailuoning injection in modified collagen can improve angiogenic effects in the chorioallantoic membrane, with the ability of stimulating vessels to grow into the modified collagen. Astragalus mongholicus plus Mailuoning have a synergetic effect. The angiogenic effects of collagen matrices loading with Astragalus Mongholicus-Mailuoning injection are by way of stimulating proliferation, migration and tube formation of thoracic aorta endothelial cells. One of the possible mechanism may through upgrade the protein expression of VEGF.