Therapeutic Mechanism Of AGGF1 Protein On Vascular Injury

Vascular injury is the central link in vascular diseases such as atherosclerosis.The role of repair strategies for vascular injury in the treatment of this type of vascular disease is self-evident.Therefore,the angiogenic factor AGGF1 is a potent angiogenic factor and is of great significance in the treatment of vascular injury.AGGF1 is a disease-causing gene found in the study of KTS disease,and studies have shown that it is similar to vascular endothelial growth factor.Both VSMC cells and EC cells have higher levels of AGGF1 expression and exhibit potent pro-angiogenic effects.In studies of AGGF1 knockout mice,it was found to be critical for angiogenesis and vascular development,proving to be an important regulator of vascular integrity.Through experimental studies on zebrafish,it was demonstrated that AGGF1 can regulate angiogenesis by activating the AKT pathway.In previous studies,AGGF1 was found to stimulate EC cell division,adhesion and migration,and the functional domain of this effect was found.In recent experiments,it has been found that during neointimal formation,proliferation of VSMC cells in carotid arteries increases after vascular injury;AGGF1protein blocks by inhibiting proliferation of VSMC in mice and promoting phenotypic transition to contractile phenotype Neointima formation after vascular injury.Its contractile phenotype is characterized by high expression of contractile proteins,typical actin proteins are alpha-SMA,SM22 and MYH11.Therefore,in order to better explain the relationship between structure and function of AGGF1,this paper mainly studies the minimal functional domain of AGGF1 protein promoting angiogenesis,and verifies its therapeutic effect by in vivo experiments in animals.To further explore key domains with vascular injury treatment,we purified a series of N-terminal and C-terminally truncated AGGF1 proteins.It is known that AGGF1 plays an important role in neointimal formation after vascular injury,and can effectively promote adhesion,division and proliferation of vascular endothelial cells EC and vascular smooth muscle cells VSMC.AKT plays an important role in regulating cell growth,proliferation,metabolism,transcription,and protein synthesis.Therefore,after vascular injury,the angiogenesis effect can be judged based on the level of phosphorylated AKT.The protein was purified by appropriate conditions,and each truncated protein was stimulated to vascular endothelial cells,and the expression of phosphorylated AKT was detected by WB.Experiments have shown that the region between amino acid 604 and amino acid 614 can effectively increase the level of AKT phosphorylation,demonstrating that the region of angiogenesis of AGGF1 protein is located in the amino acid region 604-614.To further verify this result,we performed in vivo experiments to verify.Both ?-SMA,SM22 and MYH11 are involved in the maintenance of cell movement and cell structural integrity,and the effects of angiogenesis and therapy can be judged by the level of the three.Carotid artery ligation was performed on C57BL/6J mice.The expression of?-SMA,SM22 and MYH11 was detected by QPCR technique to verify the effect of AGGF1 and truncated protein on vascular injury in animals.The experimental results showed that the in vivo experiment was consistent with the AKT phosphorylation in the cells,and the region in which the AGGF1 protein promotes angiogenesis is located in the amino acid region 604-614.In this paper,the minimal functional domain of AGGF1 protein promoting angiogenesis was studied,and the results were verified by in vivo experiments in animals for the first time,which provided ideas for the future use of AGGF1 fragments to produce angiogenic peptide drugs.