LncRNA H19 Promotes Angiogenesis Through H19/miR-454-3p/wnk1 Axis

Ischemic heart disease(IHD)is a common cardiovascular disease that seriously threatens the life and health of human beings,especially the elderly.The morbidity and mortality of IHD rank high among all diseases in the world.Revascularization is an important therapy for the treatment of severe IHD.The existing clinical treatment methods are coronary artery bypass grafting(CABG)and percutaneous transluminal coronary angioplasty(PTCA).However,because the two methods mentioned above have strict restrictions on the therapeutic time window and suitable crowd,and the treatment success rate for elderly patients is lower,which may cause more adverse outcomes.Angiogenesis is a treatment that uses a variety of methods to stimulate the formation of new blood vessels in the ischemic myocardial tissue,thereby improving the blood supply and prognosis of the ischemic area.Therefore,finding a specific new target has become the key to promote angiogenesis of ischemic myocardium in aging organisms.Long non-coding RNA(Lnc RNA)is a type of non-coding RNA,whose length is greater than 200 bp.It is widely involved in the regulation of various physiological and pathological processes and constitutes a huge regulatory network.It plays important regulatory roles in many processes such as embryonic development,aging,metabolism,organogenesis,tumorigenesis and development.Lnc RNA mainly through the mechanism of competitive endogenous RNA(competing endogenous RNA,ce RNA)(It is known that mi RNA binding to target m RNA can inhibit the translation,degrade and break the m RNA,thus down-regulating the expression of target genes.If a certain lnc RNA can also bind to the mi RNA,then the lnc RNA can compete with m RNA for the same mi RNA,resulting in reduced binding of the mi RNA and m RNA,thus preventing the mi RNA from silencing the target m RNA and indirectly upregulating the expression of the target gene)to exert its functions.Lnc RNA H19 is a gene that mainly exists in the cytoplasm and functions as a regulatory RNA or ribose regulator.It is located on human chromosome 11p15.5.H19 has a variety of biological functions,such as participating in cell proliferation,differentiation and metabolism,etc.H19 is highly expressed in various tissues of the human tissues,especially in the endothelial cells of the cardiovascular system.Studies have found that H19 can enhance the functions of human umbilical vein endothelial cells(HUVEC,Human Umbilical Vein Endothelial Cells)to form in vitro tubular structures;mouse hindlimb ischemia models can induce upregulated expression of H19,and H19 inhibitors can slow the growth of HUVEC,and Induces its accumulation in the G1 phase of the cell cycle;H19 can promote the proliferation,migration and tube formation of glioma-related endothelial cells,and enhance the functions of glioma cells to invade and angiogenesis.Many studies have shown that H19 can promote tendons healing in mice.After H19 was delivered by nanoscale vesicles,wound healing in diabetic rats was significantly accelerated,epithelial regeneration ability,maturation degree of new blood vessels and blood supply recovery were significantly improved compared with control group.However,there is no relevant research on H19 promoting ischemic myocardial angiogenesis.In addition,H19 has also been proven to reduce hypoxia-induced cardiomyocyte damage.In this study,cell experiments confirmed that knockdown of Lnc RNA H19 can inhibit the proliferation,migration and tube formation of HUVEC in vitro;overexpression of Lnc RNA H19 can promote the proliferation,migration and tube formation of HUVEC in vitro.Therefore,we used bioinformatics analysis tools(Ensembl/mi RBase database,RNAhybrid/miranda software)to predict the downstream mi RNA of Lnc RNA H19,and predicted that mi R-454-3p might be the downstream target of H19.To further explore the downstream molecular mechanism of H19 promoting angiogenesis,we predicted the downstream m RNA of mi R-454-3p,and screened a gene that might interact with H19 by ce RNA mechanism(the number of shared mi RNAs is greater than or equal to 3,hypergeometric test p<0.01),which is WNK Lysine Deficient Protein Kinase 1(wnk1)-a protein has been proven to promote angiogenesis before.After further regulating the expression of H19 by sh RNA and mimics and inhibitors of mi R-454-3p,the regulatory correlation between Lnc RNA H19,mi R-454-3p and m RNA wnk1 was confirmed by q PCR and Western blot technology.And the dual luciferase experiment further confirmed the interaction between Lnc RNA H19 and mi R-454-3p,wnk1 and mi R-454-3p,which means H19 acts as an endogenous competitive RNA for mi R-454-3p and regulated its downstream m RNA wnk1 indirectly.Finally,we established a Lnc RNA-mi RNA-m RNA regulatory pathway,which is H19-mi R-454-3p-wnk1 axis.In summary,this study has drawn the following conclusions: Lnc RNA H19 could competitively bind with mi RNA-454-3p and promote WNK1 expression through the ce RNA mechanism,thus providing a potential new therapeutic target for promoting angiogenesis and improving ischemic myocardial microcirculation in aging organisms.