Study On Effect Of IGF-1 On Injured Vascular Endothelial Cell And Mechanism Of Action

Objective:To observe the effect of insulin-like growth factor-1 (IGF-1) on proliferation and survive of injured vascular endothelial cell and preliminarily discussed the mechanism. Established the cultivated model of schwann cells, neurons, endothelial cells and made the foundation for peripheral nerve damage research.Methods:10ng/ml,20ng/ml,40ng/ml,80ng/ml,160ng/ml IGF-1 were cultured the vascular endothelial cells in vitro for 48h, growth rate was detected by MTT method. The effect of IGF-1 on vascular endothelial cell was measured by cell migration experiment, PI stain and ELISA. Schwann cells and neurons were cultivated by tissue block stick mechanical digestion method. Using S-100, beta tubulin-â…¢and vWF to immunofluorescence identify schwann cells, neurons, endothelial cells respectively. Using NB, DMEM/F-12 mixed media, according to the different growth periods, schwann cells, neurons, endothelial cells were cultured in the culture dish medium successively, immunofluorescence examination after 7d.Results:10ng/ml,20ng/ml,40ng/ml,80ng/ml IGF-1 have different degrees effect on proliferation of vascular endothelial cell. Compared to the control group with no IGF-1, the difference of the experimental group of 40ng/ml,80ng/ml IGF-1 was statistically significant difference (P< 0.05), but the difference of the experimental group of 160ng/ml IGF-1 was not statistically significant (P> 0.05). Using 40ng/ml IGF-1 in the injured endothelial cell for 48h, the speed of growth rate was faster than the control group, PI stain demonstrated the number of dead cell was fewer than the control group, the VEGF in the medium was higher than the control group, all of the differences were statistically significant (P< 0.05). Schwann cells are long spindle cell, its nucleus are ovoid or oblong. schwann cells are fish shape arrangement. The neurons cell bodies are very small. There are many apophysises growing around the cell bodies and connecting together. Endothelial cells have a spindle cells, triangle cell, clusters exist, its nucleus are clear, cytoplasm are rich, endothelial cells are "mlabstone" arrangement growth.Conclusion:10ng/ml,20ng/ml,40ng/ml,80ng/ml IGF-1 promoted the proliferation of vascular endothelial cell,160ng/ml IGF-1 inhibited the proliferation of vascular endothelial cell, prompted the low concentrations of IGF-1 promote the proliferation of vascular endothelial cell, the high concentration of IGF-1 inhibite the proliferation of vascular endothelial cell. According to the result of IGF-1 cell proliferation, further observed IGF-1 effect on proliferation and survival of injured endothelial cell. Select 40ng/ml as the experimental group, inverted microscope observed that the blank width of the experimental group was shorter than the control group's in the meanwhile phase point; DAPI/PI double stain observed that the number of dead cell in the experimental group also less than the control group's; The experimental group's VEGF was higher than the control group's, prompted that IGF-1 might promote proliferation and survive of injured vascular endothelial cell and the mechanism might be relationship with VEGF. In the laser scanning confocal microscope, schwann cells are long spindle cell, its nucleus are ovoid or oblong. schwann cells are fish shape arrangement. The neurons cell bodies are very small. There are many apophysises growing around the cell bodies and connecting together. Endothelial cells have a spindle cells, triangle cell, clusters exist, its nucleus are clear, cytoplasm are rich, endothelial cells are "mlabstone" arrangement growth.