Effect Of FHL1 Through Regulating FAK Signaling Pathway On The Pathogenesis Of Aortic Dissection

Objective:Through investigating the expression of FHL1 and the related protein of FAK signaling pathway as well as the effect of the cell function,the purpose of this research is to define the relationship of FHL1 with FAK signaling pathway in the process of regulating the aortic wall remodeling,to discuss the specific mechanism of FHL1 in pathogenesis of the aortic dissection.Method:We collected the aortas from cases of aorta dissection and 8 cases of healthy adult donors,and extracted the total protein with measuring the concentration.The expression of FHL1 and Paxillin in two groups were analyzed by Western Blotting(WB).We use Immunohistochemistry(IHC)to define the expression relativity and location of FHL1 and the related protein of FAK in the aorta and the relationship.We constructed and packaged the adenovirus regulating the expression of FHL1 to transfect the aortic vascular smooth muscle cells.The regulation effect of the adenovirus were detected by Q-RT-PCR and WB,which from the level of the mRNA and the protein respectively.We also defined the effect of the FHL1 upregulation and the FAK downregulation to the cell proliferation and the cell migration by the trial of CCK-8 and wound healing assay respectively.We constructed and packaged the adeno-associated virus downregulating the expression of FHL1 to transfect the aortic of mice through the tail vein injection.The downregulation effect of the adeno-associated virus were detected by Q-RT-PCR and WB,which from the level of the mRNA and the protein respectively.We investigated the mortality,incidence rate of dissection and the thickness of the media in aorta wall to verify the influence of FHL1 to the aorta dissection.Results:A significant down-regulation of FHL1 and Paxillin in aorta dissection was confirmed by Western Blotting.The protein of FHL1 and Paxillin expressed mainly in the cytoplasm of VSMCs in the aortic media.The adenovirus of targeted regulation FHL1 can effectively regulated the expression of FHL1 in the level of the mRNA and the protein respectively.The tail of CCK-8 confirm that upregulation the expression of FHL1 can promote cell proliferation,and this effect can be inhibited by downregulation FAK.The same result were investigate in trial of wound healing assay,downregulation the expression of FAK can restrain the effect that upregulation of FHL1 can promote the cell migration.The adeno-associated virus of Targeted regulation FHL1 can reduce the level of mRNA of FHL1 and the expression of the protein of FHL1.Downregulation the protein of FHL1 can increase the mortality the incidence rate of aorta dissection and the thickness of the media compared with the control group.Conclusions:FHL1 can play a role in the process of the vascular remodeling in the aortic dissection,related with the FAK signaling pathway.It can promote the cell proliferation and the cell migration of HASMC through the FAK signaling pathway,and influence the occurrence and development of AD.