Combined VEGF And IGF-1 Gene Transfection Mediated By Muscle Specific Adeno-associated Virus(AAV) Vector In The Treatment Of Cerebral Ischemia Via Encephalo-myo-synangiosis

Objective: In this study,skeletal muscle-specific AAV vector mediated VEGF combined with IGF-1 was constructed to treat cerebral ischemia in mice via EMS.Methods: AAV6-tMCK-GFP,AAV6-tMCK-VEGF and AAV6-tMCK-IGF-1 vectors were constructed.The transfection efficiency of the vectors was detected by transfection with AAV6-tMCK-VEGF and AAV6-tMCK-IGF-1 in mouse temporalis muscle primary myotubes under four conditions: 0%,5%,10% and 15% stretch conditions.AAV6-tMCK-GFP,AAV6-tMCK-VEGF,AAV6-tMCK-IGF-1,and combined AAV6-tMCK-VEGF and AAV6-tMCK-IGF-1 treatments were used to treat the chronic cerebral ischemia model in mice for 1 month.Middle cerebral artery occlusion was performed and the postoperative protein expression,neovascularization and cerebral infarction area were evaluated.Results: Muscle-specific vector expressed GFP protein in myotubes,but negative in other cells.AAV6-tMCK-VEGF and AAV6-tMCK-IGF-1 can stably express VEGF and IGF-1 proteins in myotubes under different stretch conditions.The combination of AAV6-tMCK-VEGF and AAV6-tMCK-IGF-1 via EMS in the treatment of cerebral ischemia mice models can significantly improve the number of capillaries and large vessels(>10?m)in the cortex and reduce the area of cerebral infarction.Conclusion: Muscle-specific vectors can specifically express proteins in the temporal muscle.The combined treatment of AAV6-tMCK-VEGF and AAV6-tMCK-IGF-1 for cerebral ischemia through EMS can improve intracranial blood supply and enhance neuroprotection.